OBJECTIVETo investigate the effect of docosahexaenoic acid (DHA) on apoptosis, migration and invasion of cervical cancer cell lines.

METHODScervical cancer cell lines Hela and Siha in logarithmic phase were treated different concentrations of DHA. The morphological changes of the cells were observed microscopically and cell apoptosis was observed using Hoechst 33258 fluorescent staining. MTT assay was used to evaluate the effect of DHA in suppressing cell growth, and flow cytometry was employed to analyze the changes of cell apoptotic rate following DHA stimulations. Wound healing assay and Transwell migration assay were used to evaluate the migration of the cell lines. The expression levels of Bax, Bcl-2 cleaved caspase3, MMP-9 and VEGF proteins were detected by Western blotting.

RESULTSDHA exposure of the cells caused obvious morphological changes and dose-dependently increased the number of apoptotic bodies in the cells. MTT assay showed that DHA inhibited the growth of the cancer cells in a time- and concentration-dependent manner. DHA also effectively suppressed migration and invasion of the cancer cells. The cells exposed to DHA showed significantly down-regulation of Bcl-2, MMP-9 and VEGF proteins and up-regulation of cleaved-caspase 3 and Bax.

CONCLUSIONDHA can promote cervical carcinoma cell apoptosis by down-regulating the anti-apoptotic proteins Bax, Bcl-2 and cleaved-caspase3 and suppress cell invasion by decreasing MMP-9 and VEGF expressions.

Apoptosis ; Caspase 3 ; metabolism ; Cell Cycle ; Cell Line, Tumor ; drug effects ; Cell Movement ; Cell Proliferation ; Docosahexaenoic Acids ; pharmacology ; Down-Regulation ; Female ; HeLa Cells ; drug effects ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Up-Regulation ; Uterine Cervical Neoplasms ; pathology ; Vascular Endothelial Growth Factor A ; metabolism ; bcl-2-Associated X Protein ; metabolism

Polycystic ovary syndrome (PCOS) is a common gynecological reproductive endocrine and metabolic disorder disease. The clinical manifestations of PCOS are diversified, with and no ovulation, insulin resistance, hyperinsulinemia and polycystic ovaries as the main pathological and physiological changes, which can lead to non-ovulation infertility and ovulatory dysfunction infertility, cause complications, such as amenorrhea, diabetes, lipid metabolism and glucose metabolism disorder, and endanger the long-term health of the body. Traditional Chinese medicine (TCM) does not have the name of this disease, and classifies PCOS to the category of "amenorrhea", "infertility" and "abdominal mass kidney" according to its clinical manifestations. Compared with western medicine, TCM has a unique view in treating the disease, and believes that the disorders of liver, kidney and spleen functions jointly affect the occurrence and development of the disease. According to the literature review, kidney-deficiency syndrome is the main syndrome type of this disease, the kidney-tonifying method has achieved a good effect in clinic. Besides, the drug use is flexible and diversified, and both primary and secondary symptoms are taken into consideration. However, in the clinical application process, the application of kidney-tonifying method in PCOS has been affected by the thinking mode of "kidney-tonifying medicine for male diseases" to some extent. In view of this, the therapeutic effect of kidney-tonifying drugs in the treatment of PCOS is preliminarily demonstrated from several aspects of the relationship between kidney deficiency and reproduction, pathogenesis of PCOS, clinical research and experimental research of PCOS, in the expectation of providing theoretical and practical supports for kidney-tonifying therapy for PCOS and other gynecological diseases.

Objective: To investigate the effect of dodder total flavone on polycystic ovary syndrome (PCOS) rat models induced by dehydroepiandrosterone (DHEA) combined with human chorionic gonadotropin (HCG). Method: Except the blank group, the remaining rats were injected with DHEA 0.06 mg·g^-1 in the morning on the nucha and 1.5 U HCG in the afternoon for 21 consecutive days. On the 16^th day after the modeling, the vaginal smear was performed to monitor the estrus cycle. Sixty rats with successful modeling were selected and divided into model group, dacin-35 group, and high, middle and low-dose dodder total flavonoids groups, with 10 rats in each group. On the day of grouping, drugs were given respectively to the drug treatment groups, and the blank group and the model group were given equal volume solvent. The drugs were given continuously for 21 days. Blood was collected from abdominal aorta 2 h later after the final administration, serum levels of testosterone (T), estrogen (E₂), luteinizing hormone (LH) and follicle stimulating hormone(FSH) were measured; rats were put to death, and the ovaries at the same part of each rat were fixed in 10% formalin solution. htoxylin eosin (HE) staining was performed, and the morphological changes in the ovaries were observed by light microscopy; the same part of the ovary was taken, and androgen receptor (AR), B-cell lymphoma-2(Bcl-2) and Bcl-2-associated X protein(Bax) were measured in the ovary by immunohistochemistry to observe the expressions of relevant proteins in the ovary; the hypothalamus and pituitary were taken, and the expressions of gonadotropin-releasing hormone (GnRH) in hypothalamus and gonadorelin releasing hormone receptor (GnRHR) in pituitary were detected by immunohistochemistry. Result: PCOS rat model was successfully replicated. Serum levels of T, E₂ and LH were significantly reduced, and FSH level of PCOS was significantly increased in each drug treatment group (P<0.01). The pituitary GnRHR level was increased to different degrees (P<0.05,P<0.01); AR and Bax levels of the ovary were decreased to different degrees (P<0.05,P<0.01); hypothalamus GnRH level was significantly decreased, and pathological changes in ovarian tissue of PCOS rats were obvious (P<0.01) in each drug treatment group. Conclusion: The PCOS rat model was successfully replicated. Total flavonoids of dodder may play a protective role in PCOS model rats by regulating the secretion of androgen, inhibiting the expression of ovarian apoptotic protein and impacting the hypothalamic-pituitary-ovary axis pathway.

Due to the increasing incidence of central nervous system diseases,especially the increasing incidence and mortality of stroke,brain-targeted drug delivery has attached more and more attention. Nasal administration,as one of the ways of brain-targeted administration,can effectively make the drug delivered to the brain in a targeted way after by passing the blood-brain barrier,providing a new idea for the treatment of central nervous system diseases. Therefore,it is a promising administration way. In recent years,the treatment of encephalopathy by nasal administration of traditional Chinese medicine has become a hot topic in the research of traditional Chinese medicine. Ischemic stroke is one of the most important diseases endangering human health. Nasal administration has a history of thousands of years in treatment of stroke. Modern medical research has proved that there is a subtle connection between the nasal cavity and the brain,and the complex and ingenious structure of the nasal cavity provides the possibility for drugs delivery to the brain through the nose. Drug administration through nasal cavity has obvious advantages in treatment of central nervous system diseases represented by ischemic stroke. Nasal administration is characterized by non-invasion,low infection,rapid absorption and brain targeting. The author will expound the theoretical basis of brain targeting of nasal administration from the aspects of anatomy and physiology,and summarize the transport pathway of drugs through the nose into the brain,the in vitro and in vivo experimental research basis of the " nose-brain"pathway,and the clinical nasal administration of traditional Chinese medicine to prevent cerebral ischemia. It provides a reference for better research of drugs to prevent and treat cerebral ischemia injury through the " nose-brain"pathway and lays a foundation for further research of the " nose-brain" pathway.
Administration, Intranasal ; Brain ; Brain Ischemia/drug therapy* ; Drug Delivery Systems ; Humans ; Medicine, Chinese Traditional ; Nasal Mucosa ; Pharmaceutical Preparations

Esophageal cancer is a digestive tract malignancy with high morbidity and mortality and mainly occurs in males. The 5-year survival rate is lower than 20%. In China， the morbidity and mortality of esophageal cancer rank the first in the world， seriously threatening national health. The pathogenesis of esophageal cancer is diverse， which is generally considered as the consequence of environmental-genetic-gene interaction. In addition to genetic factors and regional characteristics， gene mutation， RNA interference， DNA damage repair， tumor microenvironment， dietary habit， chronic adverse stimulation， and inflammatory reaction are all involved in the occurrence and development of esophageal cancer. However， there is no unified and accurate conclusion. Clarifying the exact pathogenesis of esophageal cancer is of great significance for its early screening， diagnosis， prevention， treatment， and prognosis. Surgery， radiotherapy， and chemotherapy are the three effective methods for the treatment of esophageal cancer. However， due to the atypical early symptoms， most patients have missed the best operation period when diagnosed， resulting in poor clinical prognosis. Moreover， radiotherapy and chemotherapy will cause side effects such as loss of appetite， low immune function， esophagitis， pneumonia， and malnutrition， which is not conducive to the prognosis and treatment maintenance of patients. With definite efficacies on esophageal cancer， traditional Chinese medicine （TCM）， which is flexible and diverse in the treatment， can primarily or alternatively be involved in the treatment of esophageal cancer. TCM can eliminate postoperative complications and postoperative infections and relieve adverse gastrointestinal reactions， weakened immune function， and organ damage caused by radiotherapy and chemotherapy. It can enhance clinical efficacy and improve the quality of life of patients. Therefore， it is necessary to systematically summarize the clear pathogenesis or risk factors of esophageal cancer and review the clinical characteristics of TCM in the prevention and treatment of esophageal cancer to facilitate the early screening， diagnosis， and treatment of esophageal cancer and promote the application of TCM in the prevention and treatment of esophageal cancer and related adverse reactions.

Objective: To analyze the medication rules of traditional Chinese medicine (TCM) in treating type 2 diabetes mellitus (T2DM),and explore the mechanism of action.Method: The articlesabout TCM treatment for T2DM in CNKI and Wanfang Med Online were retrieved.The data was entered into Excel 2013 to set up a database,and the factor analysis,association rules and cluster analysis were used to analyze core medication combinations by SPSS Statistics 19.0 and SPSS Modeler 14.1.kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed using BATMAN-TCM to reveal the mechanism of action.Result: A total of 92 prescriptions were included,the high frequency medicines were Astragali Radix.The major properties of TCM were cold and plain.The major flavors of TCM were sweet and bitter.The major meridian tropism of TCM was livermeridian.These high frequency medicines were deficiency-nourishing herbs,and blood circulation-invigorating and stasis-eliminating herbs.Totally 6 core medication combinations were mined through association rules.Seven common factors were extracted from factor analysis.Clustering analysis was used to get 6 kinds of herbs.KEGG pathway analysis revealed 24 biological pathways and 6 diseases relating to T2DM.Conclusion: The most commonly used Astragali Radix for treating T2DM.In clinical,it can be combined with Poria,Salviae Miltiorrhizae Radix et Rhizoma and Rehmanniae Radix for treating T2DM.At the same time,it can be combined with such herbs as deficiency-nourishing herbs,and blood circulation-invigorating and stasis-eliminating herbs.The mechanism of action of high-frequency herbs may be related to the targets of exophosphatase/phosphodiesterase (PDEs),adenosine A1 receptor gene 1(ADORA1),adenosine A2A receptor gene (ADORA2A),neuropetide second receptor (NPY2R),and endothelial carbon monoxide synthase (NOS3) by interfering with PI3K/Akt,starch and sucrose metabolism signaling pathways.

Decoction pieces are the main body of traditional Chinese medicine (TCM) clinical medication. With the modernization of TCM medication, TCM formula granules have gradually developed and begun to take shape. Although there are processing forms of TCM injection and oral liquid, they only account for a small part of TCM market and are mainly taken orally. However, when oral administration of lipid soluble ingredients is conducted, the dissolution is limited and the absorption is relatively small. In addition, it has side effects of stimulating gastrointestinal tract, resulting in low tolerance of patients. Although there are topical preparations made by water extraction and alcohol extraction, most of them are crude preparations with random dosage, which results in the waste of lipid soluble components to a certain extent. Most of the lipid soluble components of TCM have good biological activities, such as schisandrin b, tanshinone, methyl chlorogenic acid, etc., but there is no suitable medicinal form. To make the best use of "medicine", it is necessary to systematically study the lipid soluble components of TCM. On the basis of fairly mature extraction, separation and collection processes, a medicinal form with definite pharmacological effect and beneficial to its absorption is developed based on lipid soluble components. TCM essential oil has anti-aging, anti-dementia, anti-oxidation and other activities, which has been widely used in clinical, cosmeceuticals, health products and other fields. However, the effect of single essential oil is limited. Compatibility can not only enhance the efficacy and reduce toxicity, but also expand the application range of essential oil. Therefore, formula essential oil can be used as a new form of TCM, and mainly for external use, supplemented by internal use, to make up for the disadvantages of crude preparations and random dosing for external use of TCM, as well as the disadvantages of orally absorbed less and stimulated gastrointestinal tract, to meet the needs of clinical medication, faster, more accurate, better and stronger play the effect of lipid soluble components.

Endoplasmic reticulum （ER） is an important organelle responsible for protein， steroid， lipid and carbohydrate synthesis and calcium-dependent signal transduction in eukaryotic cells. ER homeostasis is essential for normal cell function. ER homeostasis imbalance can induce ER stress （ERS）， which participates in the occurrence and development of diseases of the digestive system， respiratory system， circulatory system， nervous system， reproductive system， and endocrine system， and affects body health. Among various diseases， cancers seriously endanger people′s health due to its high mortality rate， disability rate， and recurrence rate. Due to the survival characteristics of unlimited proliferation， tumor cells are often exposed to various internal and external stimuli such as hypoxia， ischemia， excessive proliferation， and starvation， which destroy intracellular protein balance and induce ERS to some extent for survival. ERS plays a major role in various tumors and has dual functions in the survival of tumor cells： promoting the survival of tumor cells by activating a series of adaptive responses， while inducing ERS-related apoptosis pathways， so as to promote tumor cell death and inhibit tumor growth and invasion. As multiple functions of ERS in tumors are reported， many scholars have tried to intervene in the progress of tumors from the perspective of ERS. The therapeutic effect of traditional Chinese medicine （TCM） on tumors has been widely recognized. TCM can participate in the regulation of tumors from many aspects， including ERS， chemoradiotherapy resistance， gastrointestinal adverse reactions caused by chemotherapy， postoperative recurrence and metastasis. Since there are few reports on the antitumor effect of TCM from the perspective of ERS， this paper expounds the influence of ERS on tumorigenesis and development and the progress of TCM intervention in tumor through ERS， in order to provide a new direction for tumor treatment.

ObjectiveDisruption of epithelial layer may instantaneously induce the generation of endogenous electric fields, which was proved to play an important role in guiding the cell migration and promoting wound healing. PIEZO1 is a kind of mechanic sensitive channel, may be regulated by voltage, is proved to involve in chemotactic migration of cells and play an important role in the process of wound healing. In this paper, the role of PIEZO1 and its downstream proteins FAK and integrin β1 in the electric field guided cell migration were investigated by HaCaT cells (human immortalized keratinocyte). MethodsCell migration was tracked by Living Cell Imaging System in directed current (DC) electric field (EF). Inhibitors and RNAi techniques were applied to study the function of PIEZO1 and other related proteins in electric fields. Western blot was used to detect the expression and phosphorylation levels of integrin β1 and FAK in electric field guided migration under EF stimulation. ResultsPiezo1 RNAi as well as Ruthenium red and GsMTx4 treatment all significantly inhibited the electrotaxis of HaCaT cells. Electric field stimulation with GsMTx4 treatment alone increased FAK phosphorylation level and the expression of integrin β1. Electric field promoted the expression level of integrin β1 and the phosphorylation level of FAK. Inhibiting the expression of PIEZO1 by RNAi significantly attenuated the phosphorylation level of FAK under EF stimulation. Inhibition of integrin β1 and FAK by inhibitor significantly decrease the electric field guided cell migration. ConclusionPIEZO1 as well as integrin β1 and FAK are involved in the electric field guided cell migration of HaCaT cells. Electric field signals regulate the expression of integrin β1 and the activation of FAK through PIEZO1-mediated signal pathway to orchestrate cell migration.

A variety of chromatographic methods were applied to research the chemical composition of the roots of Euphorbia ebracteolata. Meanwhile, the structures of these compounds were identified by spectroscopic data, such as 1D, 2D NMR and HR-ESI-MS. In the present research, a new phenolic acid compound, 1,3,4,5-tetrahydroxy-2-O-β-D-[(6'-Gallic acyl)-glucopyranyl] isopentane(1), was isolated from the roots of E. ebracteolata, as well as three known compounds, isopropyl-2-O-β-D-(6'-gallic acyl)-glucopyranoside(2), ellagic acid(3) and 3,4'-di-O-methylellagic acid(4) respectively. Additionally, compounds 2 and 4 were isolated from the Euphorbia genus for the first time.