Preparation and characterization of nanoparticles shelled with chitosan for oral insulin delivery

Nanoparticles (NPs) composed of chitosan (CS) and poly(gamma-glutamic acid) (gamma-PGA) were prepared by a simple ionic-gelation method for oral insulin delivery. Fourier transform infrared (FT-IR) spectra indicated that CS and gamma-PGA were ionized at pH 2.5-6.6, while X-ray diffractograms demonstrated that the crystal structure of CS was disrupted after it was combined with gamma-PGA. The diameters of the prepared NPs were in the range of 110-150 nm with a negative or positive surface charge, depending on the relative concentrations of CS to gamma-PGA used. The NPs with a positive surface charge (or shelled with CS) could transiently open the tight junctions between Caco-2 cells and thus increased the paracellular permeability. After loading of insulin, the NPs remained spherical and the insulin release profiles were significantly affected by their stability in distinct pH environments. The in vivo results clearly indicated that the insulin-loaded NPs could effectively reduce the blood glucose level in a diabetic rat model.     

Effects of CDB-4022 on Leydig cell function in adult male rats

CDB-4022, an indenopryridine, suppresses spermatogenesis and decreases inhibin secretion in adult male rats. In the present study, we investigated the effects of CDB-4022 on Leydig cell function. A single oral dose of CDB-4022 (2.5 mg/kg) resulted in a 2-fold decrease in serum testosterone levels after 7 days that was paralleled by a decrease in Cyp17a1 mRNA and protein levels and 17alpha hydroxylase enzymatic activity compared with vehicle-treated rats. Consistent with the lower serum testosterone levels, pituitary Lhb and Fshb mRNA levels were increased 3.2- and 2.3-fold, respectively, by CDB-4022 treatment. Ultrastructural analysis of pituitary gonadotrophs showed distended endoplasmic reticulum (ER) and fewer secretory granules in CDB-4022-treated rats, characteristic of enhanced secretory activity. Conversely, CDB-4022 increased serum progesterone levels, testicular Star mRNA and protein expression, and the number of Leydig cells per testis. Serum inhibin B levels were undetectable in CDB-4022-treated rats, while serum activin A levels were similar to controls, indicating that the CDB-4022-treated rats have an elevated activin A:inhibin B ratio. In the presence of hCG stimulation, activin A directly suppressed testosterone secretion but enhanced progesterone secretion from rat Leydig cell primary cultures. Likewise, treatment of MA-10 cells with activin A was found to enhance cAMP-stimulated progesterone secretion and STAR expression. Together, our data indicate that CDB-4022 treatment inhibits CYP17A1 and stimulates STAR expression, thereby decreasing testosterone but increasing progesterone production. We propose that unopposed actions of activin A most likely contribute to the steroid profile in rats after CDB-4022 treatment. Our findings establish CDB-4022 as a new model to examine intratesticular control mechanisms that modulate Leydig cell gene expression and function.     

A Supplement to the Chloranthaceae of China

In the present paper the longstanding confusion of three uncertain species of Chloranthus, C. monostachys R. Br., C. pernyanus Solms-Laub. and C. kiangsiensis Metcalf is clarified. In addition to the new record of C. nervosus Coll. et Hemsl. to the flora of China, the distributional ranges of Sarcandra hainanensis (Pei) Swamy et Bailey and Hedyosmum orientale Merr. et Chun are also found to be much bigger than previously known.     

福建漳州水仙花的染色体数目及命名研究

福建漳州的水仙花分为单瓣水仙及重瓣水仙两类,李时珍早就指出它们乃一物二种^[1]。李懋学等(1980)观察了福建漳州、浙江舟山、江苏崇明岛等地的水仙花的染色体数目,均为2n=30,因此认为这三个地区的水仙花都是中国水仙Narcissus tazetta L.var.chinensis Roem.,并从染色体组型分析,阐明它们全是同源三倍体^[8]。我们分别观察了漳州的单瓣水仙及重瓣水仙染色体数目,与他们观察的结果有所不同。说明漳州水仙花的染色体数目及其分类问题,有进一步研究之必要。     

A new classification scheme for the family Araliaceae

The present paper deals with the following three aspects: 1. It attempts to discuss the problems on primitive forms of the family Araliaceae. The genus Tupidanthus Hook. f. & Thoms. was considered by H. Harms (1894) and H. L. Li (1942) as primitive, whilst another genus Plerandra A. Gray was regarded as primitive by R. H. Eyde & C. C. Tseng in 1971. Having made a detailed comparison of the taxonomical characters of these two genera, the present authors believe that both genera are not the most primitive in the Araliaceae. Their affinit yis not close enough and they possibly evolved in parallel lines from a common ancestor which is so far unknown yet. 2. By studying the systems of the past, the present authors believe that none of them is entirely satisfactory. Bentham (1867) recognized five ‘series’ (in fact, equivalent to ‘tribe’ with the ending-eae of names) based on the petaline arrangement in the bud, the numbers of stamen and the types of endospem. This is a plausible fundamental treatment for the Araliaceae, but choosing the endosperm as a criteria in dividing tribe is artifical. As we know today, both ruminate and uniform endosperm are usually presente in the same genus. Seemann’s system (1868) divided the Hederaceae (excl. Trib. Aralieae) into five tribes, in addition to the locules of ovary. The criteria are essentially the same as Bentham’s. The system of Hams (1894) divided the family into three tribes. Two tribes, Aralieae and Mackinlayeae, of Bentham are retained, but other groups were combined in the Trib. Schefflereae. However, Harms did not retain one of those three oldest legitimate names which had named by Bentham, that is contrary to the law of priority in the International Code of Botanical Nomenelature. Hutchinson (1967) adopted seven tribes for the family. The criteria essentially follow those of Bentham, but the inflorescence is overstressed. The inflorescence is an artifical taxonomical character in dividing tribes, because of some dioecious plants, such as Meryta sinclairii (Hook. f.) Seem., have two types of inflorescence in male and female plants. According to Hutchinson’s arrangement, the male and female plants would be put in separate tribes. 3. The present authors are of the opinion that in the study of a natural classification of plant groups emphasis should be laid not only on the characters of the reproductive organs, but on those of vegetative organs as well. The present revised system is based principally upon the characters of both flowers and leaves of the five tribes as follows: Trib. 1. Plerandreae Benth. emend. Hoo & Tseng Trib. 2. Tetraplasandreae Hoo & Tseng Trib. 3. Mackinlayeae Benth. Trib. 4. Aralieae Benth. Trib. 5. Panaceae Benth. emend. Hoo & Tseng     

Role of M-line proteins in sarcomeric titin assembly during cardiac myofibrillogenesis

A rat polyclonal anti-M-line protein antiserum and three mouse monoclonal anti-titin antibodies (E2, F3, and A12) were used to study the spatiotemporal relationship between M-line proteins and titin during myofibril assembly in cultured chicken cardiomyocytes by immunofluorescence microscopy. In day 2 cultures, M-line proteins and titin were detected as punctate staining in most cardiomyocytes, which possessed many nonstriated fibrils. At a late stage (day 3 cultures), M-line proteins were incorporated into dot-like structures along nonstriated fibrils, while titin staining was continuous on these structures. As development progressed, M-line proteins were registered in periodic pattern in the mid-A band. In cardiomyocytes from day 5 cultures, the titin bands were separated by an unstained region, and achieved their adult doublet pattern. Thus, the organization of titin in the sarcomere appears to occur later than that of M-line proteins in the M-line. Our morphological data indicate that the early registration of M-line proteins in primitive myofibrils may guide titin filament alignment via interaction between M-line proteins and titin. In order to investigate the role of M-line proteins in the assembly of titin filaments, anti-M-line protein or anti-titin antibodies were introduced into cultured cardiomyocytes by electroporation to functionally bind the respective proteins, and the profile of myofibril assembly was examined. Cardiomyocytes from day 2–3 cultures with incorporated anti-M-line protein antibodies became shrunk, and exhibited defective myofibrillar assembly, as shown by the failure of titin to assemble into a typical sarcomeric pattern. Incorporation of anti-titin antibody E2, which recognizes the M-line end domain of titin, resulted in the failure of M-line proteins organized into the M-line structure, as shown by random, sporadic staining with anti-M-line protein antibody. These studies confirm the essential role of M-line proteins in the organization of titin filaments in the sarcomere and that the interaction between titin and M-line proteins is crucial to the formation of the M-line structure. J. Cell. Biochem. 71:82–95, 1998. © 1998 Wiley-Liss, Inc.