Hochu-ekki-to Treatment Improves Reproductive and Immune Modulation in the Stress-Induced Rat Model of Polycystic Ovarian Syndrome.

The traditional herbal medicine, Hochu-ekki-to, has been shown to have preventive effects on viral infection and stress. This study aimed to evaluate the clinical effects of Hochu-ekki-to on two stress-related rat models of polycystic ovarian syndrome. Female Sprague-Dawley rats were divided into control and treatment groups, the latter of which were subjected to stress induced by exposure to adrenocorticotropic hormone (ACTH) or cold temperatures. After these stress inductions, rats were orally treated with dissolved Hochu-ekki-to once per day for 7 days. Rats subjected to the two different stressors exhibited upregulation of steroid hormone receptors (in ovaries) and reproductive hormones (in blood), and consequent stimulation of abnormal follicle development accompanied by elevation of Hsp 90 expression (in ovaries). Treatment with Hochu-ekki-to for 7 days after stress induction increased immune functions, reduced the stress-induced activation of Hsp 90, and normalized the levels of the tested steroid hormone receptors and reproductive hormones. Our findings suggest that stress stimulations may promote the activation of Hsp 90 via the dysregulation of steroid hormone receptors and reproductive hormones, but that post-stress treatment with Hochu-ekki-to improves reproductive and immune functions in the ovaries of stressed rats.

Stress-induced activation of ovarian heat shock protein 90 in a rat model of polycystic ovary syndrome.

AIM: Polycystic ovarian syndrome is the most common endocrine disorder affecting infertile women of reproductive age. This study evaluated the activation of heat shock protein 90 (Hsp 90) during the formation of stress-induced polycystic ovaries. MATERIAL & METHODS: Female Sprague-Dawley rats (180-200 g) were subjected to one of two stress-inducing conditions; animals were either treated with adrenocorticotropic hormone daily for 18 days or were exposed to daily cold stress for three weeks. Non-treated rats sampled during proestrus or diestrous served as controls. Blood samples were collected from the left ventricles of anesthetized rats and concentrations of follicle-stimulating hormone, luteinizing hormone, estradiol, testosterone and corticosterone were measured in all rats. The expression of messenger RNA for androgen receptor, estrogen receptor-α and -ß, nerve growth factor receptor, and glucocorticoid receptor, and protein expression for Hsp 90 was also assessed in the rat ovaries. RESULTS: Stress increased glucocorticoid receptor and androgen receptor expression, and decreased estrogen expression. Nerve growth factor receptor expression was greater in treated than diestrous rats and less in treated than proestrous rats. Ovarian Hsp 90 protein expression was increased in rats treated with adrenocorticotropic hormone or cold stress. Serum follicle-stimulating hormone levels were reduced and testosterone and corticosterone levels increased by stress, whilst luteinizing hormone and estradiol levels were similar to levels in diestrous and proestrus control rats respectively. CONCLUSIONS: The results indicate that stress, via the activation of ovarian Hsp 90 and changes in steroid hormone receptor expression and serum reproductive hormone levels, may be involved in the induction of polycystic ovaries in rats.

Effects of fraction obtained from Korean Corni Fructus extracts causing anti-proliferation and p53-dependent apoptosis in A549 lung cancer cells.

Corni Fructus has traditionally been used as herbal medicine for the treatment of tuberculosis, asthma, hepatitis, and chronic nephritis in Korea, Japan, and China. This research was carried out to evaluate the proliferative-inhibitory effect of CF extracts against cancer cells and to identify the new pro-substance from medicinal plants. Among these herbal extracts extracted from KCF (Korean Corni Fructus), JCF (Japanese Corni Fructus) and CCF (Chinese Corni Fructus), KCF extracts strongly induced anti-proliferation of cancer cells in a dose-dependent manner compared with other extracts. Moreover, after treatment with CM/F3 (fraction 3 obtained from KCF extracts) for 24 h, A549 cells were evaluated by several indicators such as cell viability, LDH release, DNA fragmentation, nuclear condensation, and apoptotic proteins in vitro. CM/F3 showed the tumor-selective growth inhibitory activity in a dose- and time-dependent manner in A549 cells. Consistently, CM/F3 effectively induced the activation of bax, cytochrome-c, caspase-3, -8, -9, p53, and p21 causing apoptosis, and caused the suppression of Cdk2, pRb, and E2F1 related to cell arrest in A549 cells. These results demonstrate that CM/F3 caused not only anti-proliferation but also cell death involving cell arrest through interaction between apoptotic proteins and the upregulation of p53 in A549 cells.

Analysis of parasitic diseases diagnosed by tissue biopsy specimens at KyungHee Medical Center (1984-2005) in Seoul, Korea.

We analyzed parasitic diseases diagnosed by tissue biopsy specimens at KyungHee Medical Center (KMC) from 1984 to 2005. The total number of parasite infection cases was 150 (0.07%) out of the total 211,859 biopsy specimens submitted for histopathological examinations. They consisted of 62 cysticercosis, 23 sparganosis, 16 paragonimiasis, 15 amebiasis, 11 anisakiasis, 11 clonorchiasis, 3 ascariasis, 2 scabies, 2 enterobiasis, 2 trichuriasis, 1 leishmaniasis, 1 taeniasis, and 1 thelaziasis. Out of 62 cysticercosis cases, 55 were detected in subcutaneous tissues or the central nerve system. Eighteen out of 23 sparganosis cases were involved in muscular and subcutaneous tissues. In most anisakiasis cases, the involved organ was the stomach. The lung and the pleura were the most common site of paragonimiasis. The incidence of parasitic diseases during the first 5 years (1984-1988) was the highest of all observed periods. After 1989, similar incidences were shown throughout the period. Whereas cysticercosis was diagnosed in 34 cases during 1984-1988, no case has been diagnosed since 2000. In the case of sparganosis, the chronological incidence was almost uniform throughout the period 1984-2005. Paragonimiasis showed a similar tendency to cysticercosis. In gender and age distribution of parasitic diseases, men showed higher incidence rates than females, and the age groups of the 40s or older indicated higher infection frequencies than other age groups. Therefore, these results are a significant report to appear the tendency of human parasitic disease diagnosed by tissue biopsy in association with parasitosis at KMC in Seoul.

A case of probable mixed-infection with Clonorchis sinensis and Fasciola sp.: CT and parasitological findings.

We report here a human case probably mixed-infected with Clonorchis sinensis and Fasciola sp. who was diagnosed by computed tomography (CT) scan, serological findings, and/or fecal examination. The patient was a 43-year-old Korean female and was admitted to Kyung Hee University Hospital with the complaints of fever and abdominal pain. On admission, marked eosinophilia was noted in her peripheral blood. CT scan showed specific lesions for clonorchiasis and fascioliasis in the liver, along with lesions suggestive of amebic abscess. Micro-ELISA revealed positive results for the 2 helminthic infections. Eggs of C. sinensis and trophozoites of Entamoeba histolytica were observed in the stool. Treatment with praziquantel followed by metronidazole and tinidazole reduced abnormalities in the liver and eosinophilia. This is the first case report of a possible co-infection with 2 kinds of liver flukes in the Republic of Korea.

Radix clematidis extract inhibits UVB-induced MMP expression by suppressing the NF-kappaB pathway in human dermal fibroblasts.

Ultraviolet (UV)B irradiation induces the production of matrix metalloproteinases (MMPs), which are responsible for the degradation of collagenous extracellular matrix in connective tissues, causing skin photoaging. Although Radix clematidis is commonly used in Chinese medicine for the treatment of arthralgia, the anti-skin photoaging effects of Radix clematidis have not yet been reported. In the present study, we investigated the inhibitory effects of Radix clematidis extract (RCE) on MMP-1 and -3 expression of human dermal fibroblast cells via various in vitro experiments and elucidated the pathways of inhibition. Western blot analysis and real-time PCR revealed RCE inhibited UVB-induced MMP-1 and -3 expressions in a dose-dependent manner. UVB strongly activated nuclear factor-kappaB (NF-kappaB) activity, which was determined by IkappaBalpha degradation, nuclear localization of p50 and p65 subunit, and NF-kappaB binding activity. However, UVB-induced NF-kappaB activation was completely blocked by RCE pretreatment. These findings suggest that RCE prevents UVB-induced MMP expression through inhibition of NF-kappaB activation. In conclusion, RCE is a potential agent for the prevention and treatment of skin photoaging.

An outbreak of gnathostomiasis among Korean emigrants in Myanmar.

Thirty-eight (designated as cases) of 60 Korean emigrants who consumed raw fresh water fish in Yangon, Myanmar developed migratory swellings and creeping eruptions on the back, abdomen, flank, and other cutaneous areas 1-10 weeks later. The symptoms included itching, nodule formation, fatigue, urticaria, fever, pain on the skin, and erythematous plaques. Skin biopsies of two cases revealed no parasites. However, the mean +/- SD peripheral blood eosinophilia among the cases was 6.3 +/- 6.5% (n = 29) and 9.0 +/- 9.8% (n = 26) in two examinations. An enzyme-linked immunosorbent assay of their serum samples, using Gnathostoma doloresi adult worms as the antigen, showed mean +/- SD optical densities of 0.47 +/- 0.29 (n = 28) and 0.32 +/- 0.20 (n = 30) in two examinations and 0.12 +/- 0.09 (n = 50) in healthy controls. Two advanced third-stage larvae of G. spinigerum were found in two of six catfish purchased at a local market in Yangon. The outbreak of the human infection is suggested to have been due to G. spinigerum, which is known to live out its life cycle in the Yangon area of Myanmar.

Electroacupuncture attenuates mechanical and warm allodynia through suppression of spinal glial activation in a rat model of neuropathic pain.

Neuropathic pain remains one of the most difficult clinical pain syndromes to treat. It is traditionally viewed as being mediated solely by neurons; however, glial cells have recently been implicated as powerful modulators of pain. It is known that the analgesic effects of electroacupuncture (EA) are mediated by descending pain inhibitory systems, which mainly involve spinal opioid, adrenergic, dopaminergic, serotonergic, and cholinergic receptors. However, studies investigating the suppressive effects of EA on spinal glial activation are rare. In the present study, we assessed the cumulative analgesic effects of EA on mechanical and warm allodynia in a rat model of neuropathic pain. We investigated the clinical efficacy of EA as long-term therapy and examined its effects on spinal glia, matrix metalloproteinase (MMP)-9/MMP-2, proinflammatory cytokines and serum immunoglobulin G (IgG) concentration. Rats were randomly divided into four groups as follows: the operation group (OP), operation with EA-non acupoint (EA-NA), operation with EA-ST36 acupoint (EA-ST36), and sham operation (shamOP). Following neuropathic or sham surgery, repeated EA was performed every other day after the behavioral test. On day 53 after the behavioral test, rats were perfused for immunohistochemistry and Western blot analysis to observe quantitative changes in spinal glial markers such as OX-42, astrocytic glial fibrillary acidic protein (GFAP), MMP-9/MMP-2, and proinflammatory cytokines. Allodynia and OX-42/GFAP/MMP-9/MMP-2/tumor necrosis factor (TNF)-α/interleukin (IL)-1ß activity in the EA-ST36 group was significantly reduced, compared to the OP and EA-NA groups, and IgG in EA-ST36 rats significantly increased. Our results suggest that the analgesic effect of EA may be partly mediated via inhibition of inflammation and glial activation and repeated EA stimulation may be useful for treating chronic pain clinically.

Rams genetically superior for IGF-I do not exhibit improved male reproductive traits.

Insulin-like growth factors (IGFs) play an important role in regulating normal physiology, and may be involved in the control of reproduction. The aim of this study was to define the relationship between IGF-I concentrations and reproductive performance over the breeding and non-breeding seasons in lines of New Zealand Romney rams that had been selected for low and high blood serum IGF-I concentration. Yearling rams from two selection lines (13 from the high line and 19 from the low line) were examined in July (winter), September (autumn) and November (summer) 2006 and March (spring) 2007. Scrotal circumference including the inguinal skin was recorded. Semen was collected by electroejaculation on 4 occasions over a 12-month period. Semen was evaluated according to standard procedures (volume, motility, density and morphology). Samples were collected from four animals from each group for measurements of mRNA for IGF-I and the IGF type 1 receptor (IGF 1R) in the testis, and IGF-I, IGF 1R and the insulin receptor (IR) in the liver. Blood samples were collected via jugular venipuncture for the measurement of IGF-I, insulin and testosterone. The incidences of morphologically abnormal sperm cells, the scrotal circumference and sperm motility were higher in the breeding than in non-breeding season. Seasonal changes were found in the percentage of abnormal sperm, scrotal circumference, sperm motility and sperm density, but there were no differences between lines in any reproductive parameters. IGF-I mRNA levels were higher in the high than the low line in the liver but not in the testis, whereas the opposite was found for levels of IGF 1R mRNA. mRNA levels for the insulin receptor in the liver were higher in the high line. Plasma testosterone concentrations did not differ between lines, whereas the concentrations of IGF-I and insulin were higher in the high line. The results suggest that IGF-I may be locally produced in the liver and the testis, and that selection for high IGF-I may not be associated with improved reproductive performance in rams.

[Study on the cestodes in Theragra chalcrogamma]

This study was performed to investigate the intestinal parasites (especially cestoda) in the guts of Theragra chalcogramma in Korea. The cestode larvae collected were morphologically observed and measured, and at the same time, microscopical study was done with the section slides of the cestode larvae. They were identified on the basis of the descriptions by Schmidt and Yamaguti. The results obtained in this study are as follows: The cestode larvae were identified as Khawia sinesis, Eulacistorhynchus chiloscyllius, Tentacularia coryphaenea, Pseudonybelinia odontacantha and Nybelinia lingualis. These are new species reported in Korea. The characteristics of Khawia sinesis are its scolex lacking loculi and broad, flat and fimbriate shape. Its neck is not separated from the body but a little constricted. Eulacistorhynchus chiloscyllius is characterized by fairly long and acraspedote scolex. The bulbs are long; retractor muscles are attached to bases of the bulbs. The two bothridia are oval or round in shape and lacking posterior notch. The tentacles are long and poeciloacanthus; double chainette present. Tentacularia coryphaenea is characterized by its long craspedote and subcylindrical scolex. The bothridia are separated, without free borders and spinous. The tentacles are short, slender, armed with solid hooks in spirals, similar except the base of tentacle. The tentacle sheaths are not twisted. The bulbs are ellipsoidal. Pseudonybelinia odontacantha is characterized by its craspedote scolex. The tentacles are inserted near anterior margin of bothridia and armed with hooks in spirals ascending left to right. The hooks are similar, throughout tentacle, with feeble curve and provided with a tooth-like protuberance on the ventral side. The bulbs are three times longer than its width. The posterior margins of bothridia have a pair of eversible ciliated pits; or fossettes. Nybelinia lingualis is characterized by its short and craspedote scolex. The bothridia are separated, with free boarders. The tentacles are cylindrical, armed with solid and similar hooks in quincunxes.

[Histochemical studies on Echinorhynchus gadi (Acanthocephala)]

The distribution of glycogen, polysaccharide, mucopolysaccharide, lipid and nucleic acid has been studied in Echinorhynchus gadi(Acanthocephala). The results were summarized as follows: 1. Glycogen and polysaccharide was demonstrated by Bauer PAS reaction technique and was found in fertilization membrane in ovum, central nuclear mass in acanthor and lemnisci, hypodermis in cystacanth. 2. Mucopolysaccharide was demonstrated by Mowry alcian blue staining technique and was found in outer membrane, fibrillar coat, fertilization membrane and inner membrane in acanthocephalan ova. 3. Lipid was demonstrated by Smith Nile blue stain and Lison Sudan black B staining technique and was found roughly parallel to that of polysaccharide. 4. Nucleic acid was demonstrated by Rosenbeck Feulgen reaction, Taft methylgreen-pyronin stain and Diengdoh acridine orange staining technique and found in central nuclear mass in acanthor, also, was found in lemnisci, proboscis and hypodermis in cystacanth.