[Protective effects of total saponins of Codonopsis on ulcerative colitis induced by TNBS in rats and its mechanism].

Objective: To study the protective effects and mechanisms of total saponins of Codonopsis (TSC) on ulcerative colitis in rats. Methods: Fifty male Wistar rats were randomly divided into 5 groups: control group, model group, salazosulfadiazine (SASP) positive control group (0.3 g/kg), TSC high- and low-dose experimental groups(1.2, 0.4 g/kg). UC rat model was established by trinitrobenzene sulfonic acid (TNBS)/ ethanol enema. After administration for 21 days, the rats' symptoms and signs, disease activity index (DAI), colonic mucosal injury index (CMDI) and colonic tissue morphology were observed. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), inflammatory cytokines interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor (TNF-α) in colon tissues were determined. Protein expression of nuclear nuclear transcription factor-κB (NF-κB) in colon tissues was detected. Finally, the effect of TCS therapy was evaluated. Results: Compared with the control group, the DAI and CMDI scores of the rats in the model group were increased significantly, meanwhile the colonic mucosa was seriously damaged, indicating that the model was successful. Compared with the model group, the TSC high and low dose groups could significantly reduce the DAI and CMDI score (P＜0.05) and improve the colonic mucosa form. TSC also could increase the SOD activity and decrease MDA content in colon tissues(P＜0.05), while inhibit the levels of IL-6 and TNF-α mRNA in the colon tissues and promote the expression of IL-10 mRNA (P＜0.01). At the same time, TSC reduced the expressions of NF-κB protein in the colon (P＜0.01). The TSC high-dose group was superior to the low-dose group (P＜0.05). Conclusion: TSC has significant protective effects on ulcerative colonic mucosal damage in UC rats, and there is a dose-dependent relationship; its mechanism may be related to anti-lipid peroxidation and inhibiting the NF-κB signaling pathway to regulate the release of inflammatory factors.

[Qualitative analysis of Gynostemma longipes for medicinal usage].

The Qinling-Daba Mountains area is the main producing areas of Gynostemma longipes for medicinal usage, and samples of wild whole plants in Pingli, Shaanxi Province and Qingchuan, Sichuan Province were collected. The ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS~E) was used to profile the chemical compositions and analyze the similarities and differences of G. longipes samples in these areas. Based on the accurate molecular weight and fragment information obtained from Q-TOF-MS~E, the structures of the main components were identified by combining with the mass spectra, chromatographic behaviors of reference standards and related literatures. The results showed that the components of wild G. longipes from different places among Qinling-Daba Mountains area were similar. Forty-five chemical components were identified in the whole plant of G. longipes from Pingli, Shaanxi Province, including 43 triterpenoid saponins and 2 flavonoids which contain all main peaks in its fingerprint. The main components are dammarane-type triterpenoid saponins, such asgypenoside ⅩLⅨ, gypenoside A and its malonylated product of glycosyl.

[Effects of Biejia Yugan Granule on rat with ethanol-induced hepatic fibrosis].

Objective: To investigate the prevention and treatment effect of Biejia Yugan Granule (BYG) on ethanol-induced hepatic fibrosis (EHF) rats. Methods: SD rats were randomly divided into blank control group, model group, BYG group, colchicine group and BYG low and BYG high dose groups (n=8). The EHF rat model was established by intragastric edible ethanol with a gradually increased dose. Briefly, the rats of model group, colchicine group and BYG low and high dose groups were given gavage of 5 g/(kg·d) ethanol at week 1～4, 7 g/(kg·d) ethanol at week 5～8, 9 g/(kg·d) ethanol at week 9～12 and 9.5 g/(kg·d) ethanol at week 13～24. And the other two groups were treated with equal volume water. At the same time, the corresponding drugs were administrated daily: BYG group was treated with Biejia Yugan Granules 5.55 g/kg, colchicine group was treated with colchicine 0.1 mg/kg, BYG low-dose and high-group were treated with Biejia Yugan Granules 1.85 and 5.55 g/kg respectively. The blank control group and model control group were given the same amount of purified water. On the 169th day of the experiment, the effects of BYG on the macroscopic changes of rat liver organs, the water content of liver tissue and the pathological changes of fibrosis, the content of hydroxy proline (Hyp) in liver tissue and the expression levels of α-SMA and CREB were observed. Results: BYG at the doses of 1.85 and 5.55 g/kg could significantly improve the macroscopic changes of liver and pathological changes of liver tissue fibrosis in rats with EHF, reduce the contents of water and Hyp in liver tissue, and down-regulate the expressions of α-SMA and CREB. Conclusion: BYG has obvious effect on inhibiting EHF and one of its mechanisms is down-regulate the content of CREB.

[Effect of Yuyin Ruangan Granule on TGF-ß1 expression in hepatic fibrosis rats].

OBJECTIVES: To observe the preventive and therapeutic action of Yuyin Ruangan Granule (YRG, Traditional Chinese Medicine) in hepatic fibrosis rats model and its effect on transforming growth factor-ß1 (TGF-ß1) expression. METHODS: The Wistar rats were randomly divided into 6 group (n=10), and the model of hepatic fibrosis rats was established by subcutaneous injected with carbon tetrachloride (CCL4), fed on high-fat diet and 20% ethanol for 6 weeks, to survey the effect and mechanism of YRG preventing hypatic fibrosis by detecting liver function (the activity of alanine aminotransferase(ALT), aspartate aminotransferase(AST), etc.) of liver fibrosis rats, liver fibrosis indicators (hyaluronic acid, Ⅲ procollagen, type IV collagen, laminin and hepatic pathology, etc.), and TGF-ß1 expression in liver tissue after 6 weeks treated with YRG through intragastric administration (q. d.). RESULTS: At the 7th week, fibrotic lesions appears distinctly in liver tissue of model group compared with control group (P<0.01), YRG of 6.2~28.8 g/kg could significantly decrease hepatic index, ALT and AST activities, content of hyaluronic acid(HA), Ⅲ procollagen (PCⅢ), type Ⅳ collagen(C-Ⅳ), laminin (LN) in serum, relieve liver fibrosis pathological changes and inhibit TGF-ß1 expression in fibrotic liver tissue (P<0.05, P<0.01). CONCLUSIONS: YRG has significantly preventive effects on liver fibrosis rats model, and it may be one of its mechanisms to inhibit expression of TGF-ß1.

The pharmacodynamic active components study of Tibetan medicine Gentianopsis paludosa on ulcerative colitis fibrosis.

The current study focused on the pharmacodynamic activity components of Gentianopsis paludosa against ulcerative colitis (UC) fibrosis including symptoms of intestinal diarrhea and inflammatory. Trinitro-benzene-sulfonic acid induced UC model rats were gavaged with gradient polarity extracts respectively from ethanol-extract of Gentianopsis paludosa. Masson staining and qRT-PCR methods were respectively used to assess the degree of UC fibrosis and detect the mRNA expressions of collagen I, collagen III, a-smooth muscle actin (α-SMA) and E-cadherin in colon tissue. Separated by silica gel column chromatography, further screening was conducted until active components appeared. Infrared, nuclear magnetic resonance, mass spectroscopy and ultraviolet methods were applied to confirm active components' structures. The results indicated that the expression of collagen I, collagen III and α-SMA mRNA in the colon tissues of acetidin group rats was obviously depressed compared with control groups while E-cadherin displayed just opposite. Dyed in blue indicating UC fibrosis degree, the area of acetidin group was less than that other experimental groups. Four components: (1,8-Dihydroxy-3,7-Dimethoxyxanthones, 1-hydroxy-3,7,8-Trimethoxyxanthones, 1,7-Dihydroxy-3,8-Dimethoxyxanthones and 1-hydroxy-3,7-Dimethoxyxanthones), were obtained from acetidin group and all of which have a significant equivalence to Gentianopsis paludosa on the therapeutic effect of UC fibrosis. Our findings revealed the activity components for clinical application history of Gentianopsis paludosa and provided a preliminary foundation for further new drug research and exploitation.

[Effects of VOR on asthmatic BALB/c mice and on immune-activity of Th17 cells].

OBJECTIVE: To study the anti-asthmatic effects of Volatile oil of Radix Angelicae Sinensis (VOR, Traditional Chinese Medicine) on asthmatic BALB/c mice and its effect on Th17 cell immuno-activity through IL-17A and RORγt. METHODS: After grouping (n=12), the asthmatic BALB/c mice were replicated through injection of ovalbumin (OVA) for sensitization and administration of OVA aerosol for challenge and then, the asthmatic behaviors, respiratory function, lung histopathology as well as levels of IL-17A in serum and retinoid-related orphan receptor gamma t (RORγt) in lung tissue were observed after the action of VOR. RESULTS: VOR could maintain normal growth of body weight in asthmatic mice, improve asthmatic behaviors, respiratory function, lung histopathology and, inhibit over-expression of IL-17A and RORγt (P<0.05, 0.01) at 60, 120, 240 mg/kg doses. The combination of VOR and dexamethasone could bring synergistic effects on growth of body weight and expression of IL-17A and RORγt. CONCLUSIONS: VOR has significant effects of anti-asthma and one of the mechanisms is to inhibit immune activity of Th17 cell through relieving over-expression of IL-17 and RORγt. Besides, the combination of VOR and glucocorticoid could bring synergistic effects.

[Effect of Volatile Oil of Radix Angelicae Sinensis on experimental asthma in rats].

OBJECTIVE: To study the effects of Volatile Oil of Radix Angelicae Sinensis (VOA) on experimental asthma in rat model based on abnormal immune functions of Treg cells. METHODS: After grouping, the asthmatic rats were developed through injecting OVA and AI(OH)3 for sensitization and then administering OVA aerosol for challenge, and the respiratory functions, asthmatic behaviors, IL-10 levels in bronchoalveolar lavage fluid (BALF) (ELISA) and Foxp3 expression (immunohistochemistry) in lung of asthmatic rats were observed. RESULTS: VOA at the doses of 40-160 mg/kg could improve the respiratory functions and the asthmatic behaviors, and upgrade IL-10 levels in BALF and Foxp3 expression in lung of asthmatic rats. CONCLUSION: VOA has some effects of anti-asthma and one of the mechanisms is to improving the lower immune functions of Treg cells.

Ethanol extract from portulaca oleracea L. attenuated acetaminophen-induced mice liver injury.

Acetaminophen-induced liver injury represents the most frequent cause of drug-induced liver failure in the world. Portulaca oleracea L., a widely distributed weed, has been used as a folk medicine in many countries. Previously, we reported that the ethanol extracts of Portulaca oleracea L. (PO) exhibited significant anti-hypoxic activity. In the present study, we investigated the role of PO on acetaminophen (APAP) induced hepatotoxicity. The results demonstrated that PO was an effective anti-oxidative agent, which could, to some extent, reverse APAP-induced hepatotoxicity by regulating the reactive oxygen species (ROS) in the liver of mice. At the same time, PO treatment significantly decreased mice serum levels of IL-6 and TNFα and their mRNA expression in liver tissue IL-α and TNFα play an important role during APAP-induced liver injury. Furthermore, PO inhibited APAP and TNFα-induced activation of JNK, whose activation play an important effect during APAP induced liver injury. These findings suggested that administration of PO may be an effective strategy to prevent or treat liver injury induced by APAP.

[HPLC and UPLC-MS detection of 5-HMF from rabbit ncurolymph after treated with Cornus officinalis].

OBJECTIVE: To detect 5-HMF from rabbit ncurolymph after given different dosage of Cornus officinalis via intragastric administration by HPLC and UPLC-MS. METHODS: Rabbit ncurolymph was cramped out three days after given low, medium, and high dosage of Cornus officinalis. 5-HMF from rabbit ncurolymph was detected with HPLC and UPLC-MS, respectively. RESULTS: 5-HMF from rabbit ncurolymph was detected with HPLC method only in rabbits given high dose group. Meanwhile, 5-HMF could be detected with UPLC-MS method in rabbits given medium as well as low dose group. CONCLUSION: These results suggest that 5-HMF can cross the blood brain barrier (BBB) of rabbits and enter the rabbit ncurolymph.

[Clinical study on electroacupuncture combined with acupoint sticking therapy for treatment of chronic ulcerative colitis].

OBJECTIVE: To compare the clinical efficacy on chronic ulcerative colitis between electroacupuncture (EA) combined with acupoint sticking therapy (AST) and western medicine. METHODS: Ninety cases were randomly divided into EA + AST group and western medication group, 45 cases in each one. In EA + AST group, (1) Zhongwan (CV 12), Tianshu (ST 25), Guanyuan (CV 4), Qihai (CV 6) and (2) Shangjuxu (ST 37), Pishu (BL 20), Dachangshu (BL 25), Zusanli (ST 36) were selected for EA and AST by turns. In western medication group, Mesalazine was administered orally. Clinical symptom evaluation and colonoscopic examination were conducted respectively before and after treatment, and the differences in efficacy were compared. RESULTS: In EA + AST group, the total effective rate of clinical symptoms was 91.1% (41/45) and the total effective rate of the colonoscopic improvements in mucosal lesions was 84.4% (38/45), which were all superior to those (73.3%, 33/45; 66.7%, 30/45) in medication group (both P < 0.05). CONCLUSION: The combination of electroacupuncture and acupoint sticking therapy can improve in clinical symptoms and colonoscopic mucosal lesions significantly in the patients with ulcerative colitis and the efficacy of it is superior to that of Mesalazine.

Biphasic recruitment of transcriptional repressors to the murine cytomegalovirus major immediate-early promoter during the course of infection in vivo.

Our previous studies showed that establishment of murine cytomegalovirus (MCMV) latency in vivo is associated with repression of immediate-early gene expression, deacetylation of histones bound to the major immediate-early promoter (MIEP), changes in patterns of methylation of histones, and recruitment of cellular repressors of transcription to the MIEP. Here, we have quantitatively analyzed the kinetics of changes in viral RNA expression, DNA copy number, and recruitment of repressors and activators of transcription to viral promoters during the course of infection. Our results show that changes in viral gene expression correlate with changes in recruitment of RNA polymerase and acetylated histones to viral promoters. Binding of the transcriptional repressors histone deacetylase type 2 (HDAC2), HDAC3, YY1, CBF-1/RBP-Jk, Daxx, and CIR to the MIEP and HDACs to other promoters showed a biphasic pattern: some binding was detectable prior to activation of viral gene expression, then decreased with the onset of transcription and increased again as repression of viral gene expression occurred. Potential binding sites for CBF-1/RBP-Jk and YY1 in the MIEP and for YY1 in the M100 promoter (M100P) were identified by in silico analysis. While recruitment of HDACs was not promoter specific, binding of CBF-1/RBP-Jk and YY1 was restricted to promoters with their cognate sites. Our results suggest that sequences within viral promoters may contribute to establishment of latency through recruitment of transcriptional repressors to these genes. The observation that repressors are bound to the MIEP and other promoters immediately upon infection suggests that latency may be established in some cells very early in infection.

Establishment of murine cytomegalovirus latency in vivo is associated with changes in histone modifications and recruitment of transcriptional repressors to the major immediate-early promoter.

Human cytomegalovirus (CMV) is a ubiquitous herpesvirus with the ability to establish a lifelong latent infection. The mechanism by which this occurs is not well understood. Regulation of, for example, immediate-early (IE) gene expression is thought to be a critical control point in transcriptional control of the switch between latency and reactivation. Here, we present evidence that supports previous studies showing that the majority of genomes are quiescent with respect to gene expression. To study the possible role of epigenetic factors that may be involved in repression of ie gene expression in latency, we have analyzed changes in the patterns of modifications of histones bound to the major IE promoter (MIEP) in the kidneys of acutely and latently infected mice. Our studies show that, like herpes simplex virus, murine CMV genomes become relatively enriched in histones in latent infection. There are dramatic changes in modifications of histones associated with the MIEP when latency is established: H3 and H4 become hypoacetylated and H3 is hypomethylated at lysine 4, while H3 lysine 9 is hypermethylated in latently infected mice. These changes are accompanied by a relative loss of RNA polymerase and gain of heterochromatin protein 1gamma and Yin-Yang 1 bound to the MIEP. Our studies suggest that, in the majority of cells, CMV establishes a true latent infection, defined as the lack of expression of genes associated with productive infection, and that this occurs through changes in histone modifications and recruitment of transcriptional silencing factors to the MIEP.

Effect of metal ions on the interaction between bovine serum albumin and berberine chloride extracted from a traditional Chinese Herb coptis chinensis franch.

The effect of four metal ions Cu(2+), Ni(2+), Zn(2+) and Co(2+) on the interaction between bovine serum albumin (BSA) and berberine chloride (BC) extracted from a traditional Chinese Herb coptis chinensis franch, was investigated mainly by means of UV and fluorescence spectroscopy in this paper. The four metal ions make the quenching efficacy of BC to BSA higher than that in the absence of these metal ions because of the possible transition of BSA molecular conformation caused by metal ions. It was found that the quenching mechanism is a combination of static quenching with nonradiative energy transfer. In the presence of metal ions, the apparent association constant K(A) and the number of binding sites of BC on BSA are both decreased in a range of 8-19% and 25-28%, respectively, which indicates that the metal ions decrease the binding efficacy of BC on BSA and increase the concentration of free BC simultaneously. The scheme of interaction between BC and BSA in the presence of metal ions is a strong quenching but a weak binding.

[Effects of Yangyin Shengjin Decoction on hemorheological parameters and coagulation factors in model rabbits with syndrome of excessive heat consuming body fluid and blood stasis].

OBJECTIVE: To investigate the effects of Yangyin Shengjin Decoction (YYSJD) on hemorheological parameters and coagulation factors in model rabbits with syndrome of excessive heat consuming body fluid and blood stasis. METHODS: Rabbit model with syndrome of excessive heat consuming body fluid and blood stasis was produced. The effects of YYSJD on the blood viscosity, erythrocyte sedimentation rate (ESR), hematocrit, platelet aggregation rate, prothrombin time (PT), thrombin time (TT), kaolin partial thromboplastin time (KPTT), fibrinogen (Fg), thromboxane B(2) (TXB(2)), and 6-keto-prostaglandin F(1alpha) (6-keto-PGF(1alpha)) in the model rabbits were observed. RESULTS: YYSJD decreased the whole blood viscosity and hematocrit, inhibited the platelet aggregation, prolonged PT, TT and KPTT, and reduced the content of Fg. It also regulated the balance between TXB(2) and 6-keto-PGF(1alpha). CONCLUSION: YYSJD can promote the blood circulation, adjust the blood agglutinating function, and decrease the formation of thrombus. This is one of the pharmacological mechanisms of the therapeutic method of "nourishing yin to promote blood circulation" in the theory of traditional Chinese medicine for seasonal febrile diseases.