BACKGROUND:Adipose-derived mesenchymal stem cells have the ability to self-renew and have pluripotent potential under specific conditions in vitro, which have broad application prospects in clinical practice. However, isolation and culture of adipose-derived mesenchymal stem cells stil appear to have many difficulties and shortcomings. OBJECTIVE:To isolate and culture rabbit adipose-derived mesenchymal stem cells in vitro in order to study their morphology, cellsurface markers and biological properties as wel as to investigate the osteogenic and adipogenic potentials of adipose-derived mesenchymal stem cells in vitro. METHODS:Primary adipose-derived mesenchymal stem cells were isolated from the subcutaneous adipose tissue of posterior cervical region from New Zealand white rabbits and digested by 0.1%col agenase I. The cells were passaged and amplified by the trypsin digestion. The passage 4 adipose-derived mesenchymal stem cells were induced to differentiate after exposure to adipogenic or osteogenic medium. The oil red O staining, alkaline phosphatase and alizarin red staining were used to detect the results. The cellviability was detected by the cellcounting kit 8 method to drawn the growth curve. cellsurface markers were examined using flow cytometry. RESULTS AND CONCLUSION:The adipose-derived mesenchymal stem cells isolated from the subcutaneous adipose tissue of rabbits exhibited a fusiform adherent growth in a vortex pattern, and had a strong capability of proliferation that could be passaged stably to the 10th generation. Flow cytometry results showed that the cells highly expressed CD29, CD90, CD44, but lowly expressed CD45 and CD34. After adipogenic induction, the adipose-derived mesenchymal stem cells were positive for oil red O staining;after osteogenic induction, the cells were both positive for alkaline phosphatase and alizarin red staining. These findings suggest that the adipose-derived mesenchymal stem cells were successful y isolated and cultured from the subcutaneous adipose tissue of rabbits, and these cells are pluripotent with the potential to differentiate into adipocytes and osteoblasts, which are expected to be ideal seed cells for bone tissue engineering.

Objective: To observe the effect of acupuncture plus thunder-fire moxibustion on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor-β1 (TGF-β1) in cartilage of knee osteoarthritis (KOA) rats, and to explore the mechanism of acupuncture plus thunder-fire moxibustion in the treatment of KOA. Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a blank control group, a model group and an acupuncture-moxibustion group by random digits table, 10 rats in each group. Rats in the model group and the acupuncture-moxibustion group were injected with papain in the right posterior knee joint to prepare the models. The levels of MMP-3 and TIMP-1 in rat synovium of each group were measured by enzyme-linked immunosorbent assay (ELISA) after 2 weeks of treatment. The level of TGF-β1 was determined by Motic B5 Micro-camera system. Results:The levels of MMP-3 and TIMP-1 in the cartilage of the model group were significantly higher than those in the blank control group (allP<0.01); the levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were lower than those in the model group, and the between-group differences were statistically significant (all P<0.05). The levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were higher than those in the blank control group, and the differences were statistically significant (all P<0.05). The level of TGF-β1 in cartilage tissues of the model group was significantly lower than that in the blank control group (P<0.01); the level of TGF-β1 in the acupuncture-moxibustion group was higher than that in the model group (P<0.05), but it was lower than that in the blank control group, and the between-group difference was statistically significant (P<0.05). Conclusion: Acupuncture plus thunder-fire moxibustion can effectively recover the abnormal expressions of MMP-3 and TIMP-1 in KOA model rats and somewhat up-regulate TGF-β1, which may be one of its mechanisms of acupuncture plus thunder-fire for KOA.

Objective To explore the clinical characteristics ,diagnosis, treatment and prognosis of phaeohyphomycosis. Methods Clinical data were collected, including history, physical examination, cranial and spinal imaging. Brain biopsy was performed. Data of the pathology and incubation of brain tissue were analyzed. Responsiveness to treatment was followed up. Results A previously healthy three and half years old boy was presented to our unit, with a three- month history of recurrent headache, vomiting, progressive paraplegia accompanied by urinary continence and constipation. A computed tomogram scan and magnetic resonance imaging of the brain revealed multiple lesions located in the region of the parietal - occipital lobes, periventricular area and frontal lobe, with prominent surrounding edema and irregular peripheral enhancement of the mass after the administration of contrast materials. A cerebral biopsy was performed and the pathological report was cerebral phaeohyphomycosis. The culture of the tissue and cerebrospinal fluid grew a same fungus identified as exo-phiala dermatitidis. The patient's response to therapy was poor, the parents of the boy gave up therapy, and the boy died 1 month later. Conclusions Cerebral Phaeohyphomycosis caused by Exophiala dermatitidis is rare, but the most serious form of fungus infection. Pathology and incubation of the tissue are essential for diagnosis. There is no curative therapy and the prognosis is poor.

Objective To explore sonographic manifestation of fetal malformations of cortical development.Methods From August 2012 to January 2014 three hundred and twenty-five pregnancy women referred to our institution for fetal brain MRI,which were diagnosed or suspected of central nervous system abnormalities by prenatal ultrasound examination.Results In 325 of cases,14 cases (4％) were diagnosed of malformations of cortical development.Ten eases were indicated by prenatal ultrasound,including three cases of heterotopic gray matter,six cases of microcephaly and one case of hemimegalencephaly; four cases were missed by prenatal ultrasound,including two cases of schizencephaly,one case of tuberous sclerosis,and one case of hypoplasia.Conclusions Cortical malformations can be diagnosed by prenatal ultrasonography based on typical imaging characteristics.Prenatal ultrasound combined with MRI is a powerful tool in diagnosing fetal malformations of cortical development.

To explore the relationship of mieroglia activation,motoneuronal loss in the ventral horn of spinal cord and sciatic nerve regen-eration after the sciatic nerve injury,Sprague-Dawley rats were prepared a modal of the fight sciatic nerve crash injury.The immunoreactiv-ity(-ir)ofmicroglia and number of the motoneurons inthe ventral horn of spinal cord were detected at 3 and7 days,light and electron mi-croscopic detection of sciatic nerve degeneration and regeneration were performed at 4 weeks after the nerve injury.The results showed:(1)At 3 days after the sciatic nerve injury,OX-42-irinthe ventral horn of spinal cord begantoincrease significantly(P<0.05);(2)The number ratio of motoneurons in ventral horn of spinal cord in ipsilateral to contralateral for injury decreased markedly(P<0.05),in-dicating the numbers of ipsilateral motoneuroanl survival decreased;(3)Histological assessment showed the poor regeneration of the in-jured nerves;(4)Simvastatin(an inhibitor of cholesterol biosynthesis,having potential immunomodulatory capacities)facilitated the mi-eroglial activation.the motoneuronal survival and sciatic nerve regeneration were better than non-simvastatin-treated vehicle rats.The pres-ent results suggest that mieroglia activation in the ventral horns ofthe spinal cord may play an important protective role in the nerve regener-ation after peripheral nerve injury of the rat.

The immune system is a complex system involving multiple organs,and it is vulnerable to age,gender,environment and other factors.For a variation normal physiological range,it is a great challenge to evaluate drug-induced immunotoxicity in preclinical safety study.Histomorphologic assessment of the immune system is a recognized cornerstone in the identification of immunotoxicity at present.In this paper,the principles of pathological evaluation for immune system,and pathological evaluation for important immune organs including thymus,spleen,lymph nodes are discussed briefly,so that it is intended to assist toxicity pathologists in the accurate and consistent characterization of intended and unintended drug-induced alterations of the immune system.

It was difficult to prepare traditional Chinese medicine pellets due to the adverse characteristics of the herbal extract. In this study, Danshen extract (DS) powder mixed with different proportions of microcrystalline cellulose (MCC), lactose and starch were made into pellets by extrusion-spheronization. Particle size, span, bulk density, tapping density, compressibility, Hausner ratio and angle of repose were used to evaluate the micromeritic properties of mixing powders. Feret diameter, aspect ratio, yield, density and friability were used to evaluate the properties of the pellets. The correlations between micromeritic properties of raw material powders and the formability of their pellets were analyzed by cluster analysis, principal component analysis and partial least squares regression analysis. As a result, the particle size of the powders was negatively correlated with the size, density, yield, and was positively correlated with the friability of their pellets. The span, density, compressibility and angle of repose of the powders were positively correlated with the size, density, yield, and were negatively correlated with the friability of their pellets. So there were certain correlations between the micromeritic properties of raw material powders and the properties of their pellets prepared by extrusion-spheronization. This research provided a foundation for the technology and method of traditional Chinese medicine extract pellets.
Cellulose ; chemistry ; Drug Compounding ; methods ; Drug Implants ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Excipients ; chemistry ; Lactose ; chemistry ; Medicine, Chinese Traditional ; methods ; Particle Size ; Powders ; chemistry ; Starch ; chemistry

Objective To explore the clinical characteristics and emergency treatment for type 1 diabetes(T1DM) and diabetes ketoacidosis(DKA) in children under 5 years old.Methods Twenty-one children under 5 years old with T1DM with 10 years were retrospecti-vely reviewed.The onset situation,clinical feature and treatment of DKA were analyzed.Results The cases of little children diabetes might not have typical symptoms.The positivity of islet antibody was lower.High morbidity of DKA was found in little children and DKA was often caused by infection.Conclusions Infection may be involved in the onset and progress of childhood T1DM.Emergency treatment for DKA may involve the injection of small dose insulin,correction of the disorder of water and electrolysis and regulation of acid-base.

Microbiology is one of the fundamental courses in life science and its theories and application techniques are being developed rapidly, in order to arouse the students interest and help them master better the knowledge of microbiology, we have attemped an exploration and reform in the traditional teaching methods of this subject, our study would center on multimedia teaching methods of microbiology, its content were chiefly multimedia microbiology; multimedia microbiology experiment and microbiology network teaching.

Objective To study the influence of fluoride on the expression of osteoprotegerin(OPG) mRNA and protein in suckling rat osteoblasts. Methods Osteoblasts obtained from calvarial of suckling Wistar rats were cultured in vitro in the media supplemented with NaF at a series of doses[O(control), 1,2 and 4 mg/L groups], and OPG mRNA expression and protein were evaluated by RT-PCR and ELISA methods, respectively. Results OPG mRNA expression in suckling rat osteoblasts cultured in vitro significantly increased after exposure to NaF for 48 h and 72 h(F=333.48,808.34,P<0.05). OPG mRNA expression in suckling rat osteoblasts cultured in vitro after exposure to NaF for 48 h at different doses(0.810±0.003, 0.819±0.031 and 0.870±0.044 for 1,2 and 4 mg/L groups, respectively) compared with that of control (0.800±0.040, all P<0.05). OPG mRNA expression further increased for 72 h exposure to NaF(0.933±0.047,1.031±0.051,1.240±0.062 for 1,2 and 4 mg/L, respectively), significantly higher than that of the control (0.805±0.020,all P<0.05) and corresponding groups at 48 h. NaF doses and time exposure exhibited a significant synergistic effect on OPG mRNA expression(F=2004.16, P<0.05). NaF also enhanced OPG protein expression in suckling rat osteoblasts cultured in vitro. Significant differences were observed only in 4 mg/L group(0.228±0.014,0.277±0.048) and control(0.205±0.012,0.229±0.010) at 48 h and 72 h (P<0.05). In addition, OPG protein expression at 72 h post-exposure was higher than that at 48 h,but there was no synergistic effect between concentration and time(F=1.21,P>0.05). Conclusions The results suggested that NaF could increase OPG mRNA and protein expression in suckling rat osteoblasts with a synergistic effect between the doses and exposure time.