DJ­1 is a new prognostic marker and predicts chemotherapy efficacy in colorectal cancer.

Protein/nucleic acid deglycase DJ­1 (DJ­1) is a 20­kDa conserved protein, which belongs to the DJ­1/ThiJ/Pfp â…  protein superfamily. Immunohistochemistry was performed to investigate the expression of DJ­1 in a colorectal cancer (CRC) tissue microarray containing tumor and corresponding adjacent normal tissues. In the present study, DJ­1 expression was significantly upregulated in CRC cells and tissues, compared with that in normal colon cells and adjacent normal tissues, respectively. In addition, patients with high DJ­1 expression levels had a worse overall survival (OS) compared with patients with low expression levels. Multivariate Cox regression analysis revealed that high DJ­1 expression levels was an independent prognostic factor for patients with CRC. Moreover, DJ­1 was able to regulate the PI3K/Akt/p27/cyclin E and PI3K/Akt/mTOR signaling pathways to promote CRC cell growth and metastasis in vitro and in vivo. In addition, DJ­1 regulated the NF­κB/Snail signaling pathway to induce CRC cell epithelial­mesenchymal transition to promote migration and invasion. Notably, patients receiving LFP treatment (oxaliplatin, 5­FU and tetrahydrofolate) had an increased OS compared with patients who underwent only surgery and low DJ­1 expression levels. The findings from the present study suggest that DJ­1 may serve as a promising prognostic marker and predicts chemotherapy efficacy in patients with CRC.

COE inhibits vasculogenic mimicry in hepatocellular carcinoma via suppressing Notch1 signaling.

ETHNOPHARMACOLOGICAL RELEVANCE: Vasculogenic mimicry (VM) has been suggested to be present in various malignant tumors and associated with tumor nutrition supply and metastasis, leading to poor prognosis of patients. Notch1 has been demonstrated to contribute to VM formation in hepathocellular carcinoma (HCC). Celastrus orbiculatus extract (COE), a mixture of 11 terpenoids isolated from the Chinese Herb Celastrus orbiculatus Vine, has been suggested to be effective in cancer treatment. AIM OF THE STUDY: In the current study, experiments were carried out to examine the effect of COE on VM formation and HCC tumor growth both in vitro and in vivo. MATERIALS AND METHODS: CCK-8 assay and Nikon live-work station were used to observe the viability of malignant cells treated with COE. Cell invasion was examined using Transwell. Matrigel was used to establish a 3-D culture condition for VM formation. Changes of mRNA and protein expression were examined by RT-PCR and Western Blot respectively. Tumor growth in vivo was monitored using in vivo fluorescence imaging device. PAS-CD34 dual staining and electron microscopy were used to observe VM formation. Immunohistochemical staining (IHC) was used to examine Notch1 and Hes1 expression in tumor tissues. RESULTS: Results showed that COE can inhibit HCC cells proliferation and invasion in a concentration-dependent manner. VM formation induced by TGF-ß1 was blocked by COE. In mouse xenograft model, COE inhibited tumor growth and VM formation. Both in vitro and in vivo studies showed that COE can downregulate expression of Notch1 and Hes1. CONCLUSION: The current results indicate that COE can inhibit VM formation and HCC tumor growth by downregulating Notch1 signaling. This study demonstrates that COE is superior to other anti-angiogenesis agents and can be considered as a promising candidate in HCC treatment.

Notch1 promotes vasculogenic mimicry in hepatocellular carcinoma by inducing EMT signaling.

Hypervascularity is one of the main characteristics of hepatocellular carcinoma (HCC). However, the mechanisms of angiogenesis in HCC remain controversial. In this study, we investigate the role of Notch1 in angiogenesis of HCC. We found that Notch1 expression was correlated with formation of vasculogenic mimicry (VM) and expression of biomarkers of epithelial-to-mesenchymal transition (EMT) in the tumor specimens. Two HCC cell lines, HepG2 and MHCC97-H, with low and high Notch1 expression, respectively, were used to study the mechanism of VM formation both in vitro and in vivo. It was found that MHCC97-H cells, but not HepG2 cells form VM when they grow on matrigel in vitro. HepG2 cells gained the power of forming VM when they were overexpressed with Notch1, while knockdown Notch1 expression in MHCC97-H cells led to the loss of VM forming ability of the cells. Similar results were found in in vivo study. High expression of Notch1 in HepG2 promoted xenograft growth in nude mice, with abundant VM formation in the tumor samples. Moreover, we observed Notch1 was associated with the EMT and malignant behavior of hepatocellular carcinoma by analyzing clinical specimens, models for in vitro and in vivo experiments. HepG2 presented EMT phenomenon when induced by TGF-ß1, accompanied by Notch1 activation while MHCC97-H with knockdown of Notch1 lost the responsiveness to TGF-ß1 induction. Our results suggest that Notch1 promotes HCC progression through activating EMT pathway and forming VM. Our results will guide targeting Notch1 in new drug development.

Vasculogenic mimicry in hepatocellular carcinoma contributes to portal vein invasion.

Portal vein invasion (PVI) is common in hepatocellular carcinoma (HCC) and largely contributes to tumor recurrence after radical tumor resection or liver transplantation. Vasculogenic mimicry (VM) was an independent vascular system lined with tumor cells and associated with poor prognosis of HCC. The present study was conducted to evaluate the relationship between VM and portal vein invasion. A total of 44 HCC cases receiving anatomic liver resection were included in the study and were divided into groups with and without PVI. The prevalence of VM in each group was examined by CD34-PAS dual staining. The regulatory molecules of VM formation such as Notch1, Vimentin and matrix metalloproteinases (MMPs) were investigated by immunohistochemical staining. Analysis was performed to explore the association of PVI, VM and the VM regulatory molecules. PVI was found in 40.91% (18/44) cases and VM was found in 38.64% (17/44) cases in total samples. The incidence of VM was 72.22% (13/18) in PVI group while it was 15.38% (4/26) in non-PVI group (P<0.001), VM formation was positively correlated with PVI (r=0.574, P<0.001). The VM forming regulatory molecules such as Notch1, Vimentin, MMP-2 and MMP-9 were found to be correlated with PVI in HCC patients. Taken together, our results suggested that VM formation, alone with its regulatory molecules, is the promoting factor of PVI in hepatocellular carcinoma.

<b>Inhibitory Effect of Shoseiryuto on Substance P and CGRP </b><b>Production in the Trigeminal Nerve </b> / 日本東洋医学雑誌

It has been reported that the effect of shoseiryuto, a traditional Japanese “Kampo” medicine, on allergic rhinitis depends on several mechanisms. Previously, we reported that shoseiryuto administered in an allergic rhinitis rat model, inhibited increases of substance P (SP) and calcitonin gene-related peptide (CGRP) in the nasal mucus, which in turn are involved in the exacerbation of rhinitis symptoms and allergic symptoms. In the present research, we looked into whether the secretion of SP and CGRP are directly inhibited by the administration of shoseiryuto.<br>Histamine solution 5µl (10 mg/ml) was instilled in both nostrils of healthy, male SD rats, thereby inducing allergic symptoms, and for the group to which shoseiryuto had been pre-administered, increase in the concentration of SP and CGRP in the nasal mucus was significantly inhibited. Moreover, upon investigating SP and CGRP production in the trigeminal ganglia by means of immunostaining, it was found to be similarly significantly inhibited. The above results indicate that shoseiryuto directly inhibits the secretion of SP and CGRP in the C-fibers of nasal mucosa.

<b>Analgesic and Anti-stress Effects of Yokukansan in Rats with Adjuvant Arthritis</b> / 日本東洋医学雑誌

Yokukansan (YKS), one of the traditional Japanese “Kampo” medicines, is a mixture of extract powders from seven kinds of medicinal herbs (<i>Atractylodis Lanceae Rhizoma, Hoelen, Cnidii Rhizoma, Uncariae Uncis Cum Ramulus, Angelicae Radix, Bupleuri Radix, and Glycyrrhizae Radix</i>). YKS has been administered to fragile habitus patients who show symptoms such as emotional irritability, neurosis and insomnia, and to infants who suffer from night crying and convulsions. In recent years, YKS has been reported to be effective against pain disorders such as headache and chronic pain, but the mechanism underlying these beneficial effects is still unclear. In this study, the effect of YKS on chronic inflammatory pain and stress caused by pain were investigated using rats with adjuvant arthritis.<br>Male Wistar rats were injected with complete Freund's adjuvant into the plantar surface of the right hindpaw, and then pain thresholds and stress markers were measured. The thermal pain threshold measured with the plantar test significantly decreased, and the level of salivary chromogranin A (CgA), which is used as a mental stress marker, was significantly increased in this model. The administration of YKS controlled the activation of spinal microglia involved in the expression of chronic pain, and significantly reduced a decrease in the pain threshold. Moreover, an increase in the level of salivary CgA was significantly inhibited. The authors concluded that YKS has effects in reducing chronic inflammatory pain and the stress caused by pain.

Effect of Rosmarinic Acid on Allergic Rhinitis in Rats / 日本補完代替医療学会誌

<b>Objective:</b> In general, antiallergic drugs, steroids, and autonomic drugs are administered for patients with allergic diseases, but the chronic administration of these medicines may occasionally induce side effects. As such, several complementary and alternative medicine (CAM) are used, even though their effectiveness has not been established by a significant amount of evidence. Health food, herbal medicines, herbal tea, and aromatherapy, etc., are used for the treatment of allergic rhinitis (AR) as CAM. Rosmarinic acid is a type of polyphenol, and is reported to have antiallergic, antioxydative and anti-inflammatory properties. Rosmarinic acid is included in Perilla Herbs, which are used in health foods and herbal medicines (Kampo medicines), Rosemary and Lemon balm, which are used as herbal teas and essential oils of aromatherapy. This study was performed to evaluate the effect of rosmarinic acid on AR using AR model rats.<br> <b>Methods:</b> Six-week-old male Sprague-Dawley rats were sensitized with toluene 2,4-diisocyanate (TDI) to induce AR. Rosmarinic acid (1 mg/kg i.p. or 3 mg/kg i.p.) is administrated for 21 days. On day 22, the symptom of nasal allergy was evaluated by counting the number of sneezes within 10 minutes. Substance P (SP), calcitonin gene-related peptide (CGRP) and nerve growth factor (NGF) in the nasal lavage fluids were also measured.<br> <b>Result:</b> SP and CGRP take part in the exacerbation of rhinitis symptoms, and are secreted from the nerve ending following nerve stimulation by chemical mediators such as histamine. The administration of Rosmarinic acid had no effect on NGF, but significantly suppressed increases of sneezes, SP and CGRP dose-relatedly. Conclusion: Rosmarinic acid could control the symptom of nasal allergy, and one of the mechanisms was suppressing of the secretion of SP and CGRP.<br>

INFLUENCE OF EXERCISE ON BLOOD FLUIDITY IN RAT: INFLUENCE OF TRAINING IN HIGH TEMPERATURE ENVIRONMENTS ON ERYTHROCYTE IN RAT / 体力科学

[Objective] Perspiration is almost only heat radiation mechanism under high temperature environments. And sudoriferous water is supplied from blood. Blood flow is determined by blood fluidity, blood volume and the cardiovascular system. It was reported that strong stress decreased blood fluidity.In this experiment, we investigated the relation between blood fluidity and water supply in rats loaded with forced exercise in high temperature environment.[Methods] SPF male Wistar rats weighing 250g were used. All animals were put in high temperature environment (Wet Bulb Globe Temperature; WBGT: 28°C) through whole experimental period. The rats were divided into four groups randomly; Suitable temperature environment-Exercise-Non water intake (SEN), High temperature environment-Exercise-Non water intake (HEN), High temperature environment-Exercise-Water intake (HEW) and Baseline (BL). In a group of water supply, distilled water was served before and later exercise by sonde forcibly. The blood was collected before or later of exercise and blood and erythrocyte suspension fluidity were measured.[Results] In the HEN, hydroperoxides, blood sodium, lactic acid and adrenaline increased while blood and erythrocyte suspension fluidity were decreased significantly compared with the BL. In addition, the hematocrit did not increase even if water equivalent to 4% of body weight lost it.[Conclusion] We speculate that exercise in high temperature environment decreases blood fluidity. However, the water supply in exercise that might not be sufficiently improve blood fluidity.

INFLUENCE TO BLOOD FLUIDITY BY EXERCISE IN RAT: INFLUENCE OF TRAINING IN HIGH TEMPERATURE ENVIRONMENTS / 体力科学

[Objective] Physical exercises raise more or less body temperature. A body temperature is regulated constantly generally by homeostasis mechanism. Perspiration is only heat radiation mechanism under high temperature environments. And sudoriferous water is supplied from blood. Blood flow is determined by blood fluidity, blood volume and the cardiovascular system. It was reported that strong stress decreased blood fluidity.In this experiment, we investigated the relation between blood fluidity and water supply in rats loaded with forced exercise in high temperature environment.[Methods] SPF male Wistar rats weighing 150 g were used. All animals were put in high temperature environment (Wet Bulb Globe Temperature; WBGT: 28°C) through whole experimental period. In a group of water supply, distilled water was served before and later exercise by sonde forcibly. The rats were divided into five groups randomly; Rest-Non water intake (RN), Rest-Water intake (RW), Exercise-Non water intake (EN), Exercise-Water intake (EW) and Baseline (B). The blood was collected before or later of exercise and blood fluidity or platelet aggregation was measured.[Results] In the EN, platelet aggregation, lactic acid and corticosterone increased while blood fluidity were decreased significantly compared with the RN, RW and EW. In addition, the hematocrit did not increase even if water equivalent to 8 % of body weight lost it.[Conclusion] We speculate that exercise in high temperature environment decreases blood fluidity. However, the water supply that does not completely make up for quantity of depletion in exercise may improve blood fluidity.

Technical training to prevent medical accidents caused by deep acupuncture sticking-Confirmation of the apex position of acupuncture needle using electrical acupuncture stimulation- / 全日本鍼灸学会雑誌

[Objective]It is important for students to learn in school education how to prevent medical accidents caused by deep insertion of a needle. Also, it is essential in safe acupuncture treatment that students are always reminded there are individual variations in subcutaneous structure and to confirm the position of the needle tip in the subcutaneous tissue. To recognize the acupuncture needle tip, muscle twitch induced by electrical stimulation through the acupuncture needle tip was felt with fingers of training students. <BR>[Methods]To prevent accidental pneumothorax, acupuncture training was performed in the upper and lower extremities. After the class, we conducted a questionnaire investigation with the students.<BR>[Result]As the result, students realized that the position of the needle tip which they assumed is different from the real position in subcutaneous tissue.<BR>[Discussion]Without knowing the precise position of the acupuncture needle tip, a desirable effect is not provided, and furthermore, a medical accident may be caused. It is important to confirm the position of the acupuncture needle tip with consideration to individual variation for safe and effective acupuncture treatment. <BR>[Conclusion]We concluded that the skill to prick and insert the acupuncture needle to the correct position is so important in preventing medical accidents that training to learn correct needle tip position should be given in acupuncture education.

Examination of Condition Adjustment following Acupuncture Treatment for Competitive Sailors at the National Sports Festival / 全日本鍼灸学会雑誌

[Purpose] This report examines the immediate effect of acupuncture treatment for subjective symptoms in competitive sailors.<BR>[Method] The investigation was performed in the conditioning room at the national sports festival. Acupuncture treatment was performed for lumbago (n=108), dorsal pain (n=72). Acupuncture treatment was performed only on the area where players complained of pain. The variety of chief complaints, number of chief com-plaints, and visual analog scale (VAS) score were used for the investigation. Significance of differences in VAS score was compared by t-test.<BR>[Result] There were numerous chief complaints involving lumbago, followed in orderby dorsal pain, and othercomplaints. Acupuncture treatment decreased the VAS score fbr lumbago (n=108) from 52.3±215mm to 21. 5±16.4mm. Dorsal pain (n=72) was also decreased from 48.0±18.7mm to 18.5±15.2mm. These values were both significantly different at p<0.01.<BR>[Discussion] Acupuncture treatment was effective for lumbago, and dorsal pain of competitive sailors. As a result, it was suggested that the acupuncture treatment is an effective condition adjustment methods.

"Oketsu" and Hemorheological Changes-Examination by Micro Channel Array Flow Analyzer-(the first report) / 日本東洋医学雑誌

Scientific elucidation of “oketsu (blood stasis)” states is very important to understanding the diagnostic methods of oriental medicine. We investigated the correlation of blood fluidity to oketsu states, by micro channel array flow analyzer (MC-FAN). Twenty-seven female patients were divided into three groups: a non-oketsu group, a mildly affected group, and a severely affected group according to the diagnostic criteria of their oketsu syndrome. Immediately after collecting their venous blood, whole blood passage time was measured by MC-FAN. And we compared and investigated the correlation with degree of oketsu state, diagnostic criteria and changes of whole blood passage time after the administration of ku-oketsu drugs. As a result, whole-blood passage time of the mildly affected group and the severely affected group were significantly longer than that of the non-oketsu group, while the [an] improvement of blood fluidity was observed after medication. This study confirms that reduced blood fluidity is seen using MC-FAN in oketsu states, and that ku-oketsu drugs are able to improve these states.