The effect of androgen on wool follicles and keratin production in Hetian sheep / Efeito do androgênio na produção de folículos e queratina em ovinos Hetianos

Abstract To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.

Resumo Investigar a concentração ideal de andrógenos em cultura de folículos pilosos de carneiro Hetiano e detectar os efeitos da concentração de andrógenos na proliferação e apoptose de células foliculares, por meio de imunofluorescência e de determinação quantitativa, em tempo real, da fluorescência dos níveis de expressão gênica de proteína associada à queratina. Folículos pilosos foram isolados por microdissecção, e folículos de lã e pedaços de pele foram cultivados em várias concentrações de di-hidrotestosterona (DHT) em meio de cultura. Em seguida, medições diárias de crescimento longitudinal dos folículos capilares foram obtidas usando um micrômetro microscópico. Folículos de lã cultivados de Hetianos foram corados pelo método SACPIC para revelar a estrutura do folículo piloso, enquanto fatias de pele de carneiro foram usadas para observar a proliferação celular por imunocoloração e apoptose celular por meio do método TUNEL. Em âmbito da biologia molecular, a expressão gênica da proteína associada à queratina (Kap) foi estudada usando folículos capilares cultivados por vários dias, in vitro. Os efeitos das concentrações de andrógenos no crescimento e desenvolvimento dos folículos de lã de Hetianos foram estudados experimentalmente. Ensaios de proliferação de EdU revelaram que o andrógeno promoveu a proliferação celular dentro das papilas dérmicas do folículo piloso. A detecção de apoptose por TUNEL demonstrou que o tratamento com andrógeno poderia atrasar a apoptose celular. Os resultados da reação em cadeia da polimerase transcrição reversa quantitativa (qPCR) demonstraram que os padrões de expressão gênica da proteína de enxofre Kap1.1, KIF1.2, Kap2.12 e Kap4.2 foram significativamente maiores no grupo de ovinos Hetianos de montanha. Uma concentração de androgênio de 100 nM pode promover o crescimento de células foliculares de lã de Hetianos in vitro, resultando na superexpressão de alguns genes da família Kap.

Cysticercosis in Domestic Animals and Rodents / 中国人兽共患病学报

Cysticercus cellulosae, C. bovis, and C. taiwanensis are the three species of cysticercosis of human taeniid with their adults worms Taenia solium, T. saginata saginata and T. s. asiatica respectively. C. cellulosae is prevalent in America, Europe and Asia, C. bovis in Africa, America and Asia and C. taiwanensis in Asia. The natural infection source is pig for C. cellulosae, cattle for C. bovis and pig and wild boar for C. taiwanensis. The predilection sites are muscles for C. cellulosae and C. bovis and liver for C. taiwanensis. While the prepatent periods of these three species were 60-70 days, 60-75 days and 28 days respectively. Most C. cellulosae and a few for C. bovis and C. taiwanensis survive in pig, while most C. bovis and a few for C. taiwanensis survive in cattle. In rodent, all three species have a long life. C. cellulosae and C. bovis migrate through blood and lymph vessels to the small intestinal wall and then to the body muscles while C. taiwanensis migrate through blood vessels to the small intestinal wall and then to the liver. The minimal effective dosage of praziquantel against cysticercosis should be 100 mg/kg daily for three consecutive days.

Experimental studies on small hooks preceding large hooks in the growth and development of Taenia solium metacestodes.

In the present study, we have determined the growth and development pattern of rostellar hooklets of Taenia solium cysticerci (Zhengzhou and Harbin strains) in three pigs (1 SEM and 2 L-SEM strains) 89-196 days post experimental infection. A total of 3,675 cysticerci were collected from 3 pigs, 3,007 (82%) of 3,675 cysticerci were evaginated by enzyme method. 439 (15%) evaginated cysticerci were carefully examined and measured after dehydration, staining, and mounting on microscopic slides. Among 439 cysticerci, 234 (53%) had pair rostellar hooks, 88 (20%) with unpair hooks, 60 (14%) only small (outer row) hooks, and 57 (13%) no hooks including 34 hooks were completely dropped and 23 no hooks developed. The number ranged from 10 to 17 pairs for pair hooks and 1 to 29 for unpair ones. The length and width of rostallar hooks on the scolex of cysticerci were usually larger in the pig with longer infection time. Moreover, cysticerci with pair and unpair rostellar hooks had only small hooks and no hooks were present on their scolices. However, cysticerci with only large (inner row) hooks were not found. These findings indicate that the growth and development of small hooks precedes that of the large hooks in the formation of the two-row pattern rostellar hook in Cysticercus cellulosae.

Studies on abnormality of metacestodes and adult worms of Taenia solium and Taenia saginata asiatica in rodents and pigs.

Abnormalities are not uncommon in Taenia saginata and T. solium. After examining 328 mature proglottids from 2 adult worms from two experimentally infected hamsters, 13 (4.0%) were found to have no genital pore but with numerous testes and several vas efferents; 1 (0.3%) one genital pore with one reproductive system; 12 (3.7%) one on each side with two sets of reproductive system; 17 (5.2%) two on one side with 2 sets of reproductive system; 8 (2.4%) one on one side and two on the other side with 3 sets of reproductive system; 2 (0.6%) two on each side with 4 sets of reproductive system; 4 (1.2%) three on one side with 3 sets of reproductive system, and 4 one on one side and three on the other side with 4 sets of reproductive system. Nine evaginated abnormal cysticerci of T. s. asiatica from three experimentally infected SCID mice each had two protoscoleces and a big bladder. From two experimentally infected pigs, one abnormal cysticercus was observed to have two invaginated canals each in one end. Another one had a neck-band behind the scolex and a big bladder. This paper is not only the first report of abnormality of T. solium from hamster but also the first one of abnormal cysticerci of T. s. asiatica from pigs and mice.