[Radix Angelicae Sinensis and Radix Hedysari Enhance Radiosensitivity of Human Liver Cancer Cells to ¹²C6⁺Radiation].

OBJECTIVE: To determine the effect of Radix Angelicae Sinensis and Radix Hedysari (RAS-RH) on radiosensitivity of human liver cancer H22 cells to heavy ion ¹²C6⁺and its possible mechanism. METHODS: The experiment involved a comparison of proliferation of H22 cells (detected by CCK-8 assay) between four groups: control,drug (RAS-RH),radiation,and combination (RAS-RH+radiation). H22 cells were treated with different doses of radiation alone or radiation followed by RAS-RH. The radiation enhancement effect of RAS-RH on H22 was detected by Colony forming assay. The effect of RAS-RH on the apoptosis of H22 cells was detected by flow cytometry. The influence of RAS-RH on the expression levels of related protein Survivin and Caspase-9 was investigated by Western blot. RESULTS: RAS-RH inhibited the proliferation of H22 cells,with a time and dose dependency [inhibitory concentration 20% (IC20)=(117.60±2.15) mg/L]. The survival rate of H22 cells decreased significantly with the increase of heavy ion ¹²C6⁺. The two survival curves produced by the Graph Pad Prism 5.0 software were clearly separated. The combination group demonstrated smaller shoulder area at low dosage and lower survival rate of cells compared with radiation group,with a sensitization enhancement ratio (SER) of 1.39±0.07. The combination group (100 mg/L RAS-RH+2 Gy) had higher apoptosis rate and Caspase-9 protein expression level,and lower Survivin protein expression level,compared with other 3 groups ( P<0.01). CONCLUSION: RAS-RH has radiation sensitization effect on human hepatocellular carcinoma H22 cells. The mechanism may be related to down-regulation of Survivin protein expression and up regulation of Caspase-9 protein expression.

[Effect of Astragalus membranaceus on Peripheral Blood Th17 in Patients with Allergic Rhinitis].

Objective To observe the effect of Astragalus membranaceus (AM) on expression levels of helper T cell 17 (Th17) and its related factors in patients with allergic rhinitis (AR). Methods Peripheral blood mononuclear cells (PBMCs) were sampled from 31 AR patients (recruited in the AR group) and 22 healthy subjects (recruited in the control group). PBMCs were isolated and in-vitro inter- vened by high and low dose AM injection (2 000 and 500 µg/mL) respectively for 24 h. mRNA expression levels of related orphan receptor C (RORC) were detected by real time quantitative PCR (RT-qPCR). Ex- pression levels of IL-17A and IL-22 in the supernatant were measured by ELISA. Expression levels of Th17 cells were measured by flow cytometry. Results Expression levels of Th17, RORC mRNA, IL- 17A, and IL-22 were higher in the AR group than in the control group (P <0. 01). mRNA expression levels of RORC, Th17 and its cytokines were not changed statistically in the AR group and the control group after PBMCs were intervened by low dose AM (P>0. 05). After intervened by high dose AM,mRNA expres- sion levels of RORC decreased statistically in the AR group and the control group (P <0. 05 for the AR group, P <0. 01 for the control group). Meanwhile,the expression levels of Th-17 and its cytokines de- creased in the AR group and the control group with statistical difference (P <0. 01). Conclusions Ex- cessive activation of Th17 is one of key factors-for AR. AM could further inhibit inflammation of AR and control the inflammation state of AR possibly through inhibiting the differentiation of Th17 cells and promo- ting the release of its cytokines.

Radix Angelicae Sinensis and Radix Hedysari enhance radiosensitivity of 12C6+ radiation in human liver cancer cells by modulating apoptosis protein.

OBJECTIVES: To investigate the radiosensitizing effects of Radix Angelicae Sinensis-Radix Hedysari (RAS-RH [an ultra-filtration extract]) and its underlying mechanisms in human liver cancer cells H22. METHODS: This study was conducted between September 2010 and August 2012 in the Gansu University of Traditional Chinese Medicine, Lanzhou, China. The groups were assigned as the control group, drug (RAS-RH) group, 12C6+ radiation group, and combination group. The cell counting kit-8 assay, colony formation assay, cell cycle changes, and apoptosis analysis were carried out, and survivin and casepase-9 were evaluated by reverse transcription polymerase chain reaction and Western blot analyses in the 4 groups. RESULTS: The inhibitory effect of RAS-RH on H22 cells was dependent on both concentration and time, RAS-RH was able to enhance the radiosensitivity of H22 cells by increasing cell survival fraction and radiosensitization parameters. Apoptosis and the gap2/mitosis (G2/M) phase transition induced by 12C6+ heavy ion radiation was enhanced by RAS-RH treatment. Irradiation, combined with RAS-RH, decreased survivin expression while increasing casepase-9 expression in H22 cells. CONCLUSION: The RAS-RH increased the radiosensitivity of H22 cells of 12C6+ heavy ion radiation significantly, and its possible mechanism of radiosensitization is to enhance caspase-dependent apoptosis through the down-regulation of survivin, thus, it can be used as an effective radiosensitizer.

An aryl hydrocarbon receptor ligand acts on dendritic cells and T cells to suppress the Th17 response in allergic rhinitis patients.

A predominant Th17 population is a marker of allergic rhinitis (AR). The aryl hydrocarbon receptor (AhR) exhibits strong immunomodulation potential via regulation of the differentiation of T lymphocytes and dendritic cells (DCs) after activation by its ligand, such as 2-(1'H-indole-3'-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE). The aim of this study was to analyze the effect of AhR on Th17 differentiation by investigating the action of ITE on DCs and CD4(+) T cells from patients with AR. In all, 26 AR patients and 12 healthy controls were included in this study. The expression of interleukin (IL)-1ß, IL-6, IL-10, and IL-17 in the culture supernatant and the presence of Th17 cells in CD4(+) T cells and DC-CD4(+) T-cell co-culture system were measured before and after treatment with ITE. We show that ITE significantly induced cell secretion of IL-10 and inhibited IL-1ß and IL-6 production in DCs, and promoted IL-10 production and suppressed IL-17 expression in CD4(+) T cells in vitro. It also suppressed the expansion of Th17 cells in vitro. Our work demonstrates that ITE acts on DCs and CD4(+) T cells to inhibit the Th17 response that suppresses AR; the AhR-DC-Th17 axis may be an important pathway in the treatment of AR. ITE, a nontoxic AhR ligand, attenuated the Th17 response; thus, it appears to be a promising therapeutic candidate for suppressing the inflammatory responses associated with AR.

Study on in-vivo anti-tumor activity of Verbena officinalis extract.

We investigated the anti-tumor effects of Verbena officinalis extract on H22 tumor-bearing mice and its effect on immune function. Mice model of H22 solid tumor was established, the mice were divided into five groups and administered the extract, later, tumors were removed and inhibition rates were calculated; spleens were removed and spleen indices were calculated, and the sheep red blood cell-delayed-type hypersensitivity (SRBC-DTH) and the serum hemolysin level were determined. The Verbena officinalis extract had anti-tumor effect, with the inhibition rate reaching 38.78%, it also increased the spleen index to a certain extent, in addition, the changes in DTA and HA were not obvious compared with the model group. The Verbena officinalis extract had in vivo anti-tumor effect, while causing no damage on the immune function.

[Effects of the ultra-filtration extract mixture from Hedysarum Polybotrys on human liver cells HepG2 radiosensitivity].

OBJECTIVE: To investigate the effects of the ultra-filtration extract mixture from Hedysarum Polybotrys (UEMHP) on the radiosensitivity of HepG2 cells, and to explore its possible mechanisms. METHODS: The proliferation inhibition effects of UEMHP on HepG2 cells was detected by CCK-8 assay. The colony formation assay was used for the survival fraction (SF) analysis. The distribution of the cell cycle and the apoptosis rate were detected using flow cytometry (FCM). The survivin mRNA expression level was detected using reverse transcription-PCR assay. RESULTS: The inhibition of UEMHP on HepG2 cells was time-and dose-dependent at the concentration ranging between 5 -50 mg/L (P < 0.05). The parameters of the two curve for SF (P < 0.05) showed statistical difference between the irradiation group and the UEMHP irradiation group. UEMHP could inhibit the clone formation of HepG2 cells and enhance the radiosensitivity of HepG2 cells. The results of FCM showed that UEMHP could induce G2/M phase arrest. The apoptosis rate in the UEMHP irradiation group (21.42% +/- 3.74%) was higher than that in the control group (5.35% +/- 0.41%), the only UEMHP group (10.36% +/- 1.75%), or the irradiation group (10.58% +/- 2.01%) (P < 0.01). RT-PCR showed that the survivin mRNA expression level was lower in the UEMHP irradiation group (0.31 +/- 0.02) than in the control group (0.82 +/- 0.06) and the irradiation group (0.58 +/- 0.04) respectively, showing statistical difference (P < 0.01). CONCLUSION: UEMHP can enhance the radiosensitivity of HepG2 cells, and its possible mechanisms might be correlated to down-regulating the survivin mRNA expression and promoting the apoptosis.

Behavioral conditioning of antihistamine effects in patients with allergic rhinitis.

BACKGROUND: Allergic symptoms can be induced by behavioral conditioning. However, the conditionability of antiallergic effects has not yet been studied. Thus, we investigated whether the effects of a histamine 1 (H(1)) receptor antagonist are inducible in patients suffering from house-dust mite allergy using a behavioral conditioning procedure. METHODS: During the association phase, 30 patients with allergic house-dust mite rhinitis received a novel-tasting drink once daily, followed by a standard dose of the H(1) receptor antagonist, desloratadine, on 5 consecutive days. After 9 days of drug washout, the evocation trial commenced: 10 patients received water together with an identically looking placebo pill (water group), 11 patients were re-exposed to the novel-tasting drink and received a placebo pill [conditioned stimulus (CS); CS group] and 9 patients received water and desloratadine (drug group). RESULTS: During the association phase, desloratadine treatment decreased the subjective total symptom scores, attenuated the effects of the skin prick test for histamine and reduced basophil activation ex vivo in all groups. During the evocation trial, the water group, in which subjects were not re-exposed to the gustatory stimulus, showed a reduction in subjective total symptom scores and skin prick test results, but no inhibition of basophil activation. In contrast, re-exposure to the novel-tasting drink decreased basophil activation, the skin prick test result and the subjective symptom score in the CS group to a degree that was similar to the effects of desloratadine in the drug group. CONCLUSIONS: These data show that behaviorally conditioned effects are not only able to relieve subjective rhinitis symptoms and allergic skin reactions, but also to induce changes in effector immune functions.

Quantification of DeQi sensation by visual analog scales in healthy humans after immunostimulating acupuncture treatment.

Acupuncture is the most popular component of traditional Chinese medicine in Western countries. However, the mechanisms of its effects remain unclear. The therapeutic effect of acupuncture appears when a sensation of DeQi is achieved. We previously reported that repeated, but not single acupuncture treatment affected leukocyte circulation and blood pressure in healthy young humans. The objective of this study was to quantify DeQi sensation by using visual analog scales (VASs) and, to test whether DeQi induction is an important factor for the therapeutic effects of acupuncture in the same cohort. After either acupuncture or sham-acupuncture (placebo) treatment, a questionnaire containing five individual VASs was given to subjects to evaluate their DeQi sensation, including numbness, pressure, heaviness, warmth, and radiating paraesthesia, respectively. A separate VAS to measure their levels of anxiety during the treatment was also included. Our results showed that acupuncture significantly induced higher VAS values for numbness, pressure, warmth, and radiating paraesthesia, but not for heaviness than the placebo across three treatment sessions. Additionally, acupuncture did not induce higher anxiety levels than the placebo. These data confirm that VAS is an objective and reliable way to quantify DeQi sensation and, indicate that DeQi is unique to verum acupuncture treatment. Furthermore, either acupuncture-induced therapeutic effects or DeQi sensation should not be attributed to the stress-mediated effects. In summary, the induction of DeQi in each treatment session is an important factor for the physiological outcomes of repeated acupuncture treatment, and VASs offer objective, an easy and reliable way to assess it.

Repeated acupuncture treatment affects leukocyte circulation in healthy young male subjects: a randomized single-blind two-period crossover study.

Acupuncture is the most popular component of traditional Chinese medicine in western countries, which has been widely used in the treatment of numerous medical conditions, e.g., pain, emesis or asthma. However, the effects of acupuncture on neuroendocrine and immune functions in humans remain unclear. Therefore, the present study was performed to analyse whether acupuncture treatment affects leukocyte circulation as well as plasma levels of cortisol and norepinephrine in humans. Ten healthy young male subjects were enrolled in a randomized single-blind two-period crossover study. Each period contained three sessions of either acupuncture or sham acupuncture (placebo) treatment. After randomisation, the group 1 (n=5) received acupuncture treatment at acu-points ST36, LI11, SP10, and GV14, while sham acupuncture was performed for group 2 (n=5). Two weeks later, each group received the alternative treatment. Blood samples were taken before needling, 10 min after, and 30 min after removing the needles in the first and the third session. In addition, blood pressure and heart rate were determined simultaneously. Although acupuncture treatment did not affect leukocyte circulation in peripheral blood after the first session, we observed a significant decrease in leukocyte and lymphocyte values after the third session. In contrast, cortisol and norepinephrine plasma levels remained unchanged by acupuncture. These data indicate that repeated acupuncture treatment can affect leukocyte circulation in healthy humans by still unknown mechanisms.