[Acupoint application of herbal paste relieves symptoms and naso-mucosal inflammatory response by down-regulating serum TGF-ß1 in allergic rhinitis rats].

OBJECTIVE: To observe the effect of acupoint application of herbal paste on symptoms of allergic rhinitis (AR), serum immunoglobulin E (IgE) and transforming growth factor beta 1 (TGF-ß1) level, and number of nasal eosinophils (EOS) in rats with AR, so as to explore its underlying mechanisms. METHODS: Forty male Wistar rats were randomly divided into normal control, model, medication and acupoint application groups (n＝10 rats per group). The AR model was established by intraperitoneal (i.p.) injection of mixture solution of ovalbumin, aluminum hydroxide and normal saline (once every other day, for 7 times), and nasal drip plus spray inhalation of ovalbumin (on the following day of i．p., once daily for 9 days). For acupoint application, the prepared herbal paste (containing White Mustard Seed, Rhizoma Corydalis, unprocessed Radix Kansui, Herba Asari and ginger juice) was applied to bilateral "Feishu" (BL13), "Pishu" (BL20) and "Shenshu" (BL23) for 2 h, once every other day for 7 times. The rats in the medication group were given Fluticasone Propionate nasal spray daily for 14 days. Scores of nasal itching, sneezing and nasal discharge on the day after modeling and the ending of the intervention were used to evaluate behavioral changes. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of serum IgE and TGF-ß1, and the infiltration state of EOS in the nasal mucosa tissue was observed under light microscope after HE staining. RESULTS: After modeling and compared with the normal control group, the behavioral scores and the levels of serum IgE and TGF-ß1 were significantly higher (P<0.05), and the infiltration state of EOS got worse. Compared with the model group, the increased behavioral score and serum IgE and TGF-ß1 levels were evidently suppressed (P<0.05) and EOS infiltration severity in the nasal mucosa was obviously milder in both medication and acupoint application groups. No significant differences were found between the medication and acupoint application groups in behavioral score and serum IgE and TGF-ß1 levels (P>0.05). CONCLUSION: Acupoint application can improve the symptoms of AR rats, which may be associated with its effect in down-regulating the levels of serum IgE and TGF-ß1.

In Vivo Study on Site of Action of Sinapine Thiocyanate following Acupoint Herbal Patching.

OBJECTIVE: To investigate the site of action of sinapine thiocyanate (ST), following acupoint herbal patching (AHP). METHODS: Twenty Wistar rats were randomized into five groups (groups A, B, C, D, and E), and all groups received the same AHP in vivo. Skin samples were excised at 2 h, 4 h, 6 h, 10 h, and 26 h after AHP administration from group A to group E separately and the concentrations of ST in the skin were determined using a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method. A pharmacokinetic profile of ST following AHP was performed at the same time in a group of five Wistar rats to detect plasma levels at the same time intervals. RESULTS: The mean ± SD ST concentrations (ng/ml) at 2 h (group A), 4 h (group B), 6 h (group C), 10 h (group D), and 26 h (group E) after AHP administration were 250.01 ± 61.99, 61.01 ± 30.41, 40.12 ± 26.94, 78.66 ± 59.43, and 19.55 ± 18.95, respectively. No ST was detected in rats' plasma samples at the same time points. CONCLUSIONS: The site of action of ST following AHP is in the skin.

Downregulation of human Cdc6 protein using a lentivirus RNA interference expression vector.

Eukaryotic CDC6 gene function is required for the initiation of DNA replication and is a key regulatory protein during cell cycle progression. The human CDC6 gene is not expressed in most normal tissues, in contrast with its marked expression in proliferating cancer cells. An effective way to explore the gene functions of CDC6 is to knock-down the CDC6 messenger RNA (mRNA) and examine the phenotypic consequences. In this chapter, we describe the construction of a lentivirus vector to express a CDC6 DNA segment. The transcript is able to fold by itself because the sense and antisense regions are complementary. There is a 9-nucleotide (nt) loop region allowing for the short hairpin RNA (shRNA) to form. Cellular ribonucleases process the shRNA into a functional short interfering RNA (siRNA). Down-regulation of Cdc6 protein is confirmed by Western blots.