AIMTo understand the biological functions of the ectoplasmic specializations between Sertoli cells and maturing spermatids in seminiferous epithelia.

METHODSIn order to disrupt the function of the ectoplasmic specializations, nectin-2, which is expressed at the specialization, was neutralized with anti-nectin-2 antibody micro-injected into the lumen of the mouse seminiferous tubule. Anti-nectin-3 antibody was also micro-injected into the lumen in order to neutralize nectin-3, which is expressed at the specialization.

RESULTSThe actin filaments at the specialization disappeared, and exfoliation of maturing spermatids was observed by electron microscopy.

CONCLUSIONNectin-2 was neutralized by anti-nectin-2 antibody and nectin-3 was neutralized by anti-nectin-3 antibody, respectively. Inactivated nectin-2 and nectin-3 disrupted the nectin-afadin-actin system, and finally the actin filaments disappeared. As a result, the specialization lost the holding function and detachment of spermatids was observed. One of the functions of the specialization seems to be to hold maturing spermatids until spermiation.

Actins ; metabolism ; Animals ; Antibodies ; immunology ; pharmacology ; Cell Adhesion Molecules ; immunology ; metabolism ; Cell Communication ; drug effects ; physiology ; Intercellular Junctions ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Microfilament Proteins ; metabolism ; Microscopy, Confocal ; Nectins ; Seminiferous Epithelium ; cytology ; drug effects ; metabolism ; Sertoli Cells ; cytology ; drug effects ; metabolism ; Spermatids ; cytology ; drug effects ; metabolism

AIMTo isolate and transplant germ cells from adult mouse testes for transplantation.

METHODSIn order to distinguish transplanted cells from endogenous cells of recipients, donor transgenic mice expressing green fluorescent protein (GFP) were used. Germ cells were collected from the donors at 10-12 weeks of age and spermatogonia were concentrated by percoll fractionation and transplanted into recipient seminiferous tubules that had been previously treated with busulfan at 5 weeks of age to remove the endogenous spermatogenic cells.

RESULTSTwenty weeks after the transplantation, a wide spread GFP signal was observed in the recipient seminiferous tubules. The presence of spermatogenesis and spermatozoa was confirmed in sections of 12 out of 14 testes transplanted (86 %). However, when germ cells were transplanted without concentration the success rate was zero (0/9).

CONCLUSIONGerm cells from adult mouse testes can be successfully transplanted into recipient seminiferous tubules if the cell population is rich in spermatogonia and the percoll fractionation is useful in obtaining such a cell population.

Animals ; Cell Fractionation ; Green Fluorescent Proteins ; Luminescent Proteins ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Seminiferous Tubules ; cytology ; physiology ; Spermatogenesis ; physiology ; Spermatogonia ; physiology ; transplantation ; Testis ; cytology

Fukushin, or abdominal diagnosis, is one of the characteristics of the Kampo medical examination. During the Edo period, “Fukushokiran (腹証奇覧)” became popular due to its explanations of illustrations of abdominal diagnosis and findings, focusing on the prescriptions in “Shanghanlun (傷寒論)” and “Jinguiyaolue (金匱要略).” In this study, we examined various existing editions of “Fukushokiran” and highlighted the differences in abdominal diagnosis illustrations and findings among them. According to the year indicated at the beginning and end of the volume, “Fukushokiran” is divided into pre-Kyowa and Bunka editions. Although the currently facsimile edition is based on the Bunka edition, significant differences were observed in abdominal diagnosis illustrations and other findings, which were revised from the pre-Kyowa edition. “Fukushokiran” has two parts including the first and second parts; however, differences between the pre-Kyowa and Bunka editions are particularly noticeable in the second part. Inaba Bunrei, the author of “Fukushokiran,” died in Bunka 2 (1805) ; therefore, it is unlikely that he was involved in the revision of the Bunka edition. Instead, it is assumed that the views of Wakuta Shukuko, a disciple of Bunrei, are reflected in the Bunka edition.

There are few reports on the effects for children using the diagnostic method of Kampo medicine (called sho : pattern diagnosis). Therefore, we investigated the effects of combination therapy with Kampo medicines prescribed by specialists in this ancient form of medicine to children who were refractory to modern medicine. This was a retrospective observational study of 98 children (pre-school children (PS ; n = 21), elementary school students (ES ; n = 37) and junior high school students (JS ; n = 40)) younger than 16 years old at the first visit to our department of Chiba University hospital between April 2007 and April 2017. We collected the following information from their medical records : background, chief complaint, referral source, and efficacy of Kampo medicines. We evaluated efficacy for Kampo medicine as follows : Higher improvement (HI) (symptoms improved by 2/3 or more) ; Improvement (I) (symptoms improved by 1/3-2/3) ; Mild improvement (MI) (symptoms improved by 1/3 or less) ; constant ; aggravated ; and unknown or first visit only. Positive response (HI, I, and MI) due to Kampo medicines was observed in 76% (75/98) of patients. No aggravations were observed. The chief complaint by age was skin diseases in PS, digestive diseases in ES, and cardiovascular diseases in JS. Pediatrics was the primary referral in all ages, while 30% of JS were referred from psychiatry. Kampo medicine was significantly more effective for those in PS and ES compared to those in JS (p = 0.025). Combined use of Kampo medicine and modern medicine therapy is useful for children refractory to modern medicine.

We herein report a patient who complained persistent exudate at navel in spite of adequate antibiotics therapy after laparoscopic cholecystectomy was successfully treated with Kampo therapy. The pharmacological effect of this Kampo formulation i.e. senkinnaitakusan is still unknown, but this formulation has been used for persistent infectious diseases. According to the description in the classic textbook, this formulation is suggested to encourage metabolic function and exhibit anti-inflammatory function. There have been no reports of Kampo therapy for the superficial incisional site infection associated with laparoscopic surgery. The authors propose that Kampo medicine is another promising option in the management of surgical site infection (SSI).

We herein report three patients who complained atypical febrile diseases associated with painful whole body (Case 1), general fatigue (Case 2), and a sense of general discomfort (Case 3). All cases were not accompanied by dizziness and a shaky feeling which are commonly associated in shimbuto-sho i.e. indication of shimbuto, but presented severe tenderness at the left paraumbilical region which was suggested as indicative point of shimbuto by Yoshiko Takagi. We made a diagnosis in these three cases as shimbuto-sho based on the Takagi's tender point. Through these clinical experiences we propose that a new type of shimbuto-sho, which are completely different from the fixed idea of inherited knowledge. This new type of shimbuto-sho consists of following signs ; fever with chill, floating and feeble pulse, no sweating, general fatigue or body pain, and severe pain at Takagi's tender point.

We investigated the number of drugs and pharmaceutical cost among 159 patients prescribed Western medicine and hospitalized from August 2006 to August 2015 in the Department of Oriental (Kampo) Medicine at Chiba University Hospital. The number of drugs used in Western medicine among improved patients significantly decreased from 5.6 ± 3.6 at hospitalization to 5.3 ± 3.5 at discharge, but the number of Kampo medicine drugs was not changed. The total number of drugs including both Western medicine and Kampo medicine significantly decreased from 7.0 ± 3.8 to 6.7 ± 3.6. The number of drugs used in Western medicine among nochanged patients decreased from 5.1 ± 3.4 at hospitalization to 5.0 ± 3.7 at discharge, but the number of Kampo medicine drugs significantly increased from 1.0 ± 0.0 at hospitalization to 1.3 ± 0.5. The total number of drugs including both Western medicine and Kampo medicine increased from 6.1 ± 3.4 to 6.3 ± 3.9. We thus conclude that a combination of Kampo medicine with Western medicine can be useful for reducing the number of drugs related to polypharmacy. To achieve these results, it is essential to use the concept of sho (a way of pattern recognition of a patient's symptoms in Kampo medicine).

The Koho school has a high regard for formulation corresponding to sho uniquely developed as a conspicuous feature of Kampo medicine, and Ruihobunrui (the classification of formulae based on the main crude drug : CF) has played an important role in structuring the foundation of this development in the Koho school. CF is a classification method used to express the adaptation and characteristics of formulae in the Koho school, and is useful for research on drug compositions. Therefore, CF can be used effectively in both clinical practice and education. CF has shown little development in China after the appearance of Xu Ling Tai ; however, in Japan the development of CF centered on such influential figures as Todo Yoshimasu of the Koho school, and Naohiro Kitamura of the Kosho school. Kenzo Okuda and Keisetsu Otsuka used CF for clinical practice and education during the revival stage of Kampo medicine in the Showa era ; moreover, CF had a large effect on structuring the foundation of clinical application by formulating Kampo extracts for prescriptions. Thus, CF has played an important role in Kampo medicine. However, the research has been stopped in the present circumstances of Japan. We focus here on CF of the Koho school in Edo and Showa eras, and carried out considerations of CF to establish one of the characteristics of Kampo medicine that places importance on Shang Han Za Bing Lun.

We herein report a 36-year-old male patient, who complained of persistent and recurrent nausea with vomiting for 16 years in spite of various standard treatments, was successfully treated with Kampo formulation Shimbuto, which selected based on the left para-navel tender point. We speculated that the clinical disorder in this case was related to suitai i.e. water imbalance syndrome in Kampo medicine. However, there has never been such a case report in the field of Kampo, and no related papers have been found in the standard database Medline. We believe this report is valuable from the viewpoint of therapeutics in both Western and Kampo medicine.

We experienced 8 cases of children who presented with somatic complaints and found it difficult to attend school successfully treated with Kampo medicine. We evaluated them objectively by using the evaluation scale in the guideline for medical care of school refusal. Four cases were orthostatic dysregulation, 2 cases were migraine, 1 case was involuntary movement in upper abdomen and the other case was autonomic dysfunction. Kampo medicines were selected according to conventional method, so-called sho. Among 8 cases, 5 cases had a major response, 2 cases had a minor response and one case had no beneficial response. These results suggest that Kampo therapy can be effective therapeutic means. Kampo therapy is quite personalized and beneficial especially for children who find it difficult to attend school and need individualized care. This is the first report evaluating many cases objectively by using the evaluation scale and there have been no reports similar to ours. We have to continue to enlighten general pediatricians about the effectiveness of Kampo therapy.