Tangshen formula targets the gut microbiota to treat non-alcoholic fatty liver disease in HFD mice: A 16S rRNA and non-targeted metabolomics analyses.

BACKGROUND: Tangshen formula (TSF) has an ameliorative effect on hepatic lipid metabolism in non-alcoholic fatty liver disease (NAFLD), but the role played by the gut microbiota in this process is unknown. METHOD: We conducted three batches of experiments to explore the role played by the gut microbiota: TSF administration, antibiotic treatment, and fecal microbial transplantation. NAFLD mice were induced with a high-fat diet to investigate the ameliorative effects of TSF on NAFLD features and intestinal barrier function. 16S rRNA sequencing and serum untargeted metabolomics were performed to further investigate the modulatory effects of TSF on the gut microbiota and metabolic dysregulation in the body. RESULTS: TSF ameliorated insulin resistance, hypercholesterolemia, lipid metabolism disorders, inflammation, and impairment of intestinal barrier function. 16S rRNA sequencing analysis revealed that TSF regulated the composition of the gut microbiota and increased the abundance of beneficial bacteria. Antibiotic treatment and fecal microbiota transplantation confirmed the importance of the gut microbiota in the treatment of NAFLD with TSF. Subsequently, untargeted metabolomics identified 172 differential metabolites due to the treatment of TSF. Functional predictions suggest that metabolisms of choline, glycerophospholipid, linoleic acid, alpha-linolenic acid, and arachidonic acid are the key metabolic pathways by which TSF ameliorates NAFLD and this may be influenced by the gut microbiota. CONCLUSION: TSF treats the NAFLD phenotype by remodeling the gut microbiota and improving metabolic profile, suggesting that TSF is a functional gut microbial and metabolic modulator for the treatment of NAFLD.

Sleep deprivation affects sex hormones secretion by regulating the expression of the circadian clock gene in the hypothalamus and pituitary via the PI3K/Akt signaling pathway in pregnant rats.

Sleep deprivation (SD) has many deleterious health effects and occurs in more than 70% of pregnant women. However, the changes in sex hormones and relevant mechanisms after SD have not been well clarified. The aim of the present study was to explore the effects of SD on the secretion of sex hormones and the underlying mechanisms. Twelve pregnant Wistar rats were divided into control (CON, n = 6) and SD (n = 6) groups. Pregnant rats in the SD group were deprived of sleep for 18 h, and allowed free rest for 6 h, and then the above procedures were repeated until delivery. The CON group lived in a 12 h light/dark light cycle environment. Estradiol (E2) and progesterone (P4) levels were detected by enzyme-linked immunosorbent assay (ELISA), and the expression of circadian clock genes, Bmal1, Clock and Per2, in hypothalamus and pituitary gland tissues were evaluated by immunohistochemistry (IHC) and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The PI3K and Akt phosphorylation levels in the hypothalamic and pituitary tissues were determined by Western blot. The results showed that, compared with the CON group, the SD group exhibited significantly reduced serum E2 and P4 levels, down-regulated Bmal1, Clock and Per2 expression, as well as decreased phosphorylation levels of PI3K and Akt. But there was no significant difference of the total PI3K and Akt protein expression levels between the two groups. These results suggest that SD might affect the expression of the circadian clock genes in the hypothalamus and pituitary via PI3K/Akt pathway, and subsequently regulate the secretion of sex hormones in the pregnant rats, which hints the important roles of SD-induced changes of serum sex hormone levels in the pregnant rats.

Isoliquiritigenin inhibits the proliferation, migration and metastasis of Hep3B cells via suppressing cyclin D1 and PI3K/AKT pathway.

The overall survival rate of patients with hepatocellular carcinoma (HCC) has remained unchanged over the last several decades. Therefore, novel drugs and therapies are required for HCC treatment. Isoliquiritigenin (ISL), a natural flavonoid predominantly isolated from the traditional Chinese medicine Glycyrrhizae Radix (Licorice), has a high anticancer potential and broad application value in various cancers. Here, we aimed to investigate the anticancer role of ISL in the HCC cell line Hep3B. Functional analysis revealed that ISL inhibited the proliferation of Hep3B cells by causing G1/S cell cycle arrest in vitro. Meanwhile, the inhibitory effect of ISL on proliferation was also observed in vivo. Further analysis revealed that ISL could suppress the migration and metastasis of Hep3B cells in vitro and in vivo. Mechanistic analysis revealed that ISL inhibited cyclin D1 and up-regulated the proteins P21, P27 that negatively regulate the cell cycle. Furthermore, ISL induced apoptosis while inhibiting cell cycle transition. In addition, phosphatidylinositol 3'-kinase/protein kinase B (PI3K/AKT) signal pathway was suppressed by ISL treatment, and the epithelial marker E-cadherin was up-regulated when the mesenchymal markers Vimentin and N-cadherin were down-regulated. In brief, our findings suggest that ISL could be a promising agent for preventing HCC tumorigenesis and metastasis.

Effects and Mechanism of Qiwei Qinggan Powder on Hepatic Fibrosis Based on Proteomics / 中国药房

OBJECTIVE：To investigate the anti- hepatic fibrosis （HF）effects of Qiwei qinggan powder and explore its possible mechanism. METHODS ：Male Wistar rats were randomly divided into blank group ，HF model group ，Qiwei qinggan powder low-dose，medium-dose and high-dose groups [ 135，270，405 mg/（kg·d），by total amount of crude drugs] ，with 12 rats in each group. Except for blank group ，other groups were given 50％ CCl4-peanut oil solution intragastrically （2 mL/kg，twice a week ，for consecutive 8 weeks） to induce HF model. At same time ， blank group and model group were given constant volume of 0.5％ CMC-Na solution intragastrically ；administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 8 weeks. General situation of rats were observedand liver morphology was observed after last administration and hepatic indexes were detected. The contents of liverfunction indexes （ALT，AST，ALP，HYP）in serum and the expression of α-SMA in hepatic tissue were determined ， and HE and Masson staining were performed to observe the histopathology. Using the difference multiple of expression quantity as the index ，TMT technology was used to screen the differentially expressed protein in medicine group （combining the liver tissue samples of Qiwei qinggan powder groups ）and HF model group. Uniprot-GOA database and KAAS ，KEGG mapper online tools were used to analyze GO and KEGG pathway enrichment. RESULTS ：The rats in the blank group were in good health ；the liver was bright red and smooth ，the liver lobules were intact ，no degeneration and necrosis ，inflammatory cell infiltration or fibrous tissue proliferation was found. Compared with blank group ，the rats in HF model group had poor diet ，depressed spirit ，disordered and lusterless fur ；the liver was dark red or yellow with rough surface ，hard texture ，inflammatory cell infiltration ，fiber tissue destruction ，bridge connection and so on ；the hepatic index ，the contents of liver function indexes and the expression of α-SMA were increased significantly （P＜0.05）. Compared with HF model group ，above symptoms of rats were improved to different extent in different dose groups of Qiwei qinggan powder ；hepatic index in Qiwei qinggan powder low-dose group ，the content of ALP in high-dose group ，the contents of ALT，AST and HYP and the expression of α-SMA in different dose groups were decreased significantly （P＜0.05）. A total of 42 differentially expressed proteins related to HF were screened ，of which 15 were up-regulated and 27 were down-regulated in expression，including fatty acid binding protein 4（FABP4），cholesterol 7α-hydroxylase（CYP7A1）. The results of enrichment analysis showed that the differentially expressed proteins were mainly enriched in extracellular space ，blood particles and other cell parts，involving the molecular functions of oxidoreductase activity and fatty acid binding ，the biological processes of the regulation of heterotypic cell adhesion ，protein activation cascade ，as well as retinol metabolism ，arachidonic acid metabolism ，PPAR and other signal pathway. CONCLUSIONS ：Qiwei qinggan powder can reduce the hepatic index ，ALT，AST，ALP and HYP contents in serum ，down-regulate the expression of α-SMA，improve the degree of inflammation and fibrosis of liver tissue ，and have a certain protective effect on rats. The anti-HF mechanism of it involves multiple targets and signal pathways ，such as FABP 4， CYP7A1 and PPAR.

Protective effects of Schisandrin on high glucose-induced changes of RhoA and eNOS activity in human umbilical vein endothelial cells.

Schisandrin, derived from the Chinese medicinal herb Schisandra chinensis, has been found to confer protective effects on circulation systems. But the underlying molecular mechanisms remain unclear. The aim of this study was to investigate the effects of a high level of glucose on RhoA and eNOS activity in human umbilical vein endothelial cells(HUVECs) and how Schisandrin plays a role in mediating these effects. To find the optimal treatment time, HUVECs were cultured at a high glucose concentration (30 mM) for different lengths of time (0, 12, 24, and 48 h). Subsequently, the cells were randomized into five groups: a normal group, a high glucose group, and three high glucose groups that were given different doses (5, 10, and 20 µM) of Schisandrin. The cells were pretreated with Schisandrin for 24 h before stimulation with high glucose. The morphology of HUVECs in the various groups was assessed under a light microscope. Immunocytochemical staining was used to detect the level of p-MYPT1 expression. The levels of RhoA activity were determined using the RhoA Activation Assay Biochem Kit. The levels of eNOS activity were examined using a nitrate reduction test. The results showed that in the high glucose group, the activity of RhoA was increased and the activity of eNOS was reduced, thus decreasing the secretion of NO. However, after pretreatment with Schisandrin (10, 20 µM), the activity of RhoA was inhibited and the activity of eNOS increased, which led to an increase in NO production compared with the high glucose group. There was no evident difference between the 5 µM Schisandrin group and the high glucose group. Taken together, these findings indicate that Schisandrin can improve the function of endothelial cells by lowering the activity of RhoA/Rho kinase and raising both the activity of eNOS and the production of NO.

Influence of acupuncture on expression of T-type calcium channel protein in airway smooth muscle cell in airway remodeling rats with asthma / 中国针灸

<p><b>OBJECTIVE</b>To observe the anti-remodeling effect of acupuncture on asthma and to explore the mechanism of T-type calcium channel protein in airway smooth muscle cell in airway remodeling effect in asthma.</p><p><b>METHODS</b>Thirty-two rats were randomly divided into a normal group, a model group, an acupuncture group and a sham acupuncture group, 8 rats in each group. The rats in the latter three groups were sensitized for consecutive 14 days by single peritoneal injection of aqueous suspension 1 mL of 10 mg ovalbumin (OVA), 200 mg aluminum hydroxide and saline together with 1 mL inactivated pertussis vaccine. From the 15th day, asthma was induced for 30 minutes by ultrasonic atomizing inhalation of 1% OVA for consecutive 14 days in the model group. The acupuncture group was treated with acupuncture at "Dazhui" (GV 14), "Fengmen" (BL 12) and "Feishu" (BL 13) for 30 minutes before the ultrasonic atomizing inhalation, once every two days for consecutive 14 days. The same acupoints selection and the course of treatment as the acupuncture group were produced in the sham acupuncture group and they were treated with acupuncture at 1 mm acupoint skin without retaining needles. The normal group remained unhandled. The respiratory function and the airway remodeling were evaluated by airway resistance and pulmonary histopathology, respectively, and the T-type calcium channel protein expression of Ca(v)3.1, Ca(v) 3.2, Ca, 3.3 in airway smooth muscle cell were detected by immunohistochemistry technique.</p><p><b>RESULTS</b>(1) The airway resistance in the model group was higher than that in the normal group and in the acupuncture group (both P < 0.05), and the airway resistance in the acupuncture group was lower than that in the sham acupuncture group (P < 0.05). (2) The ratios of the airway wall thickness to the basement membrane perimeter (Awt/Pbm) and the airway outer perimenter to the airway internal perimeter (Po/Pi) in the model group were higher than those in the normal group and in the acupuncture group (all P < 0.05), and the ratios of Awt/Pbm and Po/Pi in the acupuncture group were lower than those in the sham acupuncture group (both P < 0.05). (3) The average optical of Ca(v) 3.1 and Ca(v) 3.2 in airway smooth muscle cell in the model group were higher than that in the normal group and in the acupuncture group (both P < 0.05), and the average optical of Ca(v) 3.3 in airway smooth muscle cell in the model group was higher than that in the normal group (P < 0.05) and it was lower than that in the sham acupuncture group (P < 0.05).</p><p><b>CONCLUSION</b>Acupuncture can inhibit the airway remodeling and the accrementition of the airway smooth muscle and can reduce the airway resistance. The mechanism may be related to the inhibition of T-type calcium channel protein in airway smooth muscle cell, especially in relation to the protein expression of Ca(v) 3.1.</p>

Discussing the diagnosing and treating mode of corresponding prescription and pattern in Japanese kampo medicine / 国际中医中药杂志

Japanese kampo medicine originated from traditional Chinese medicine. But different from TCM pattern of "Tracing the etiology by syndrome differentiation-establishing therapeutic methods-prescribing formula", during the process of development, Japanese kampo medicine has gradually formed a different diagnostic and therapeutic mode, which was "corresponding prescription and pattern". This therapeutic mode directly chose formula from ancient books, especially Shanghanlun (Treatise on Febrile Diseases), according to the constitution, symptoms and signs of patients. The diagnostic and therapeutic mode of Japanese kampo medicine provides references for integration of traditional Chinese and western medicine.

Effects of Rukuaixiao decoction on hyperplasia of mammary gland in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of Rukuaixiao decoction (RKX) on hyperplasia of mammary gland in rats.</p><p><b>METHOD</b>60 SD female rats were radomly divided into blank control group, model group, tamoxifen group and different dose of RKX groups. Injection of estradiol and progesterone were given to establish rat models of mammary gland hyperplasia and RKX was given at the same time. Changes of breast diameter, mammilla height were measured; serum sex hormones levels, blood rheology, SOD activity and MDA content were estimated. Pathologic changes of mammary gland in rats were also observed under light microscope.</p><p><b>RESULT</b>RKX could decrease the increased breast diameter, mammilla height, reduce the numbers of mammary gland lobules and relieve the pathologic changes of mammary gland. It could also decrease estradiol, prolactin levels and MDA content in serum, increase the serum progesterone level and inhibit the decrease of the coefficient of thymus.</p><p><b>CONCLUSION</b>Rukuaixiao decoction has the function of treatment on hyperplasia of mammary gland.</p>

Pharmacognostic studies on Desmodium gyrans / 中国中药杂志

<p><b>OBJECTIVE</b>To study the characteristic features of Desmodium gyrans in order to provide a basis for rational exploitation and utilization of the herb.</p><p><b>METHOD</b>Samples of the title plant were collected, the microscopic features of cross sections and powders were studied. TLC profiles and UV absorption of the plant extract were examined.</p><p><b>RESULT</b>Calcium oxalate crystals were found in cells of transverse sections. Nonglandular hairs were observed on leaf surfaces. Characteristic peaks in the UV spectrum were identified.</p><p><b>CONCLUSION</b>The distinct characteristic features revealed in this studies can serve as evidence for the identification of D. gyrans.</p>