[Biological characteristics of sheep peripheral blood mesenchymal stem cell].

OBJECTIVE: To obtain eripheral blood mesenchymal stem cells (PBMSCs) from sheep and rabbits by continuous mobilization of granulocyte colony-stimulating factor (G-CSF), and by comparing the success rates, cell yields and biological characteristics of the two sources of PBMSCs, and to provide the experimental basis for the preclinical study of PBMSCs transplantation to repair articular cartilage injury and cartilage tissue engineering. METHODS: Through morphological characteristics, flow cytometry analysis of its surface markers, and induction of trilineage differentiation of the two cells in vitro (ie: adipogenic differentiation, osteogenic differentiation, chondrogenic differentiation), the obtained cells were finally confirmed to be PBMSCs. The colony-forming units (CFUs) and the acquisition success rates of the two PBMSCs were counted and compared, and the production of the second generation of the two PBMSCs was counted and compared by hemocytometer, and the cell counting kit-8 was used to detect the doubling time of the two PBMSCs, and the results of trilineage differentiation were quantitatively analyzed by image processing. RESULTS: Microscopically, the PBMSCs of fusiform sheep and rabbits were arranged in fish group, and the second generation of sheep and rabbit PBMSCs expressed CD44 and CD90, but not CD34 and CD45. The induction of trilineage differentiation of the two cells in vitro were successful. The CFUs of primary sheep and rabbits PBMSCs were: 7.27±1.56, 5.73±1.62, and the success rate of acquisition of sheep and rabbits PBMSCs were 78.57% and 36.67%. The number of the second-generation sheep and rabbits PBMSCs that obtained per milliliter of peripheral blood were: 29 582±2 138, 26 732±2 286, and the cell doubling times (h) of the third-generation sheep and rabbits PBMSCs were: 22.32±0.28, 33.21±0.64, the cell doubling time (h) of the fourth generation sheep and rabbits PBMSCs were: 23.62±0.56, 35.30±0.38, and the quantitative lipid ratio of sheep and rabbit PBMSCs were: 7.77%±3.81%, 17.05%±1.52%, sheep and rabbit PBMSCs chondroglobus acid mucopolysaccharide positive ratios were: 11.67%±0.53%, 8.14%±0.57%. There were statistical differences among the above groups (P < 0.05). CONCLUSION: The continuous mobilization of G-CSF to obtain sheep PBMSCs is more efficient. Sheep PBMSCs have more abundant yield and stronger proliferation ability.Sheep PBMSCs can produce more acidic mucopolysaccharides and have lower adipogenic abi-lity under appropriate conditions. Sheep PBMSCs have good research prospects in repair of articular cartilage injury with autologous stem cell transplantation and preclinical animal in vivo experiment of cartilage tissue engineering.This experiment provides further experimental basis for this kind of research.

[Epidemiological study of bone and joint injury based on urban medical insurance database].

OBJECTIVE: To estimate the prevalence rate of bone and joint injury in China and to describe the three-dimension distribution of the disease (area, time and people). METHODS: Based on a cross-sectional design, a retrospective study was conducted by using Chinese basic medical insurance database from January 1, 2013 to December 31, 2017 to analyze the epidemiological characteristics of bone and joint injury. The prevalence rate of bone and joint injury in each city was calculated, and then using meta-analyses to estimate the pooled prevalence of each area and the whole country. The pooled prevalence rates were compared among the different groups of populations, in terms of geographical area, time and population characteristics (age and gender). RESULTS: A total of 28 419 264 subjects were included in this study, including 705 793 patients with bone and joint injury. From 2013 to 2017, in Chinese basic medical insurance database, the overall prevalence rate of bone and joint injury was 141.5(95%CI: 90.4－203.7) per 10 000 population, and the prevalence rates of non-specific or polyarticular disease, knee disease, and shoulder disease were 101.6 (95%CI: 63.5－148.4)per 10 000 population, 22.5(95%CI:15.1－31.4)per 10 000 population and 10.9 (95%CI: 6.4－16.4)per 10 000 population. The prevalence rates varied across the areas, the highest rate was observed in North China, with the prevalence of 310.6 (95%CI: 12.6－989.7) per 10 000 population, and the lowest rate was observed in Southwest China, with the prevalence of 59.0 (95%CI: 37.5－85.2) per 10 000 population. The prevalence rate of bone and joint injury increased over the study period, from 111.1 (95%CI: 56.0－182.5)per 10 000 population in 2013 to 175.5 (95%CI: 116.8－245.5)per 10 000 population in 2017. The prevalence of bone and joint injury in the female population was 149.1 (95%CI: 94.2－215.9) per 10 000 population, which was higher than that of men [133.6(95%CI: 86.2－190.9) per 10 000 population]. The higher prevalence of knee disease, unspecified or polyarticular disease, and bone and joint injury were observed in people aged 60 years and older, while the prevalence of shoulder disease peaked in 40－59 years old people [20.6 (95%CI: 12.5－30.5) per 10 000 population]. CONCLUSION: This study reported a relative low prevalence of bone and joint injury in China from 2013 to 2017. The prevalence increased over the study period, and the highest prevalence rate was observed in North China. The prevalence rate showed differences among different groups of populations, and higher rates were observed in females and people aged 60 years and older.

Enhanced thermal conductivity and tensile strength of Al-17Si-3.5Cu with SiC-nanoparticle addition.

The morphology and size of primary Si has a significant influence on the thermal conductivity (TC) and strength of Al-17Si-3.5Cu. In this study, the effect of a 1-3 wt% SiC nanoparticle (SiCnps) addition on TC and tensile strength of Al-17Si-3.5Cu was investigated. Nanoparticles distributed at the interface between primary Si and Al led to a significant refinement of primary Si; for example, a primary Si size of 2 µm with 3 wt% SiCnps addition was achieved. TC of SiCnps/Al-17Si-3.5Cu improved with an increase in nanoparticle content. Nanoparticles distributed at the interface between Si and Al reduced the interfacial thermal resistance. Thus, the effective TC of eutectic Si increased. Owing to the refinement of the primary Si and the increased interfacial thermal resistance, originating from the high content of SiCnps at the interface, the effective TC of primary Si decreased. Compared with Al-17Si-3.5Cu, contribution to the improvement of the TC of SiCnps/Al-17Si-3.5Cu resulted mainly from eutectic Si. Due to the refinement of primary Si, the tensile strength of SiCnps/Al-17Si-3.5Cu improved with an increase in SiCnps content. When the SiCnps content was 3 wt%, the yield strength, ultimate tensile strength and elongation of SiCnps/Al-17Si-3.5Cu were ∼176 MPa, 418 MPa and 7%, respectively, which were improved by 37.5%, 53.7% and 218%, respectively, when compared with Al-17Si-3.5Cu.

Early infant diagnosis and outcomes in HIV-exposed infants at a central and a district hospital, Northern Malawi.

Setting: Mzuzu Central Hospital (MZCH), Mzuzu, and Chitipa District Hospital (CDH), Chitipa, Malawi. Objective: To compare management and outcomes of human immunodeficiency virus (HIV) exposed infants in early infant diagnosis (EID) programmes at MZCH, where DNA polymerase chain reaction (PCR) testing is performed on site, and CDH, where samples are sent to MZCH, between 2013 and 2014. Design: Retrospective cohort study. Results: Of infants enrolled at MZCH (n = 409) and CDH (n = 176), DNA PCR results were communicated to the children's guardians in respectively 56% and 51% of cases. The median time from sample collection to guardians receiving results was 34 days for MZCH and 56 days for CDH. In both hospitals, only half of the dried blood spot (DBS) samples were collected between 6 and 8 weeks. More guardians from MZCH than CDH received test results within 1 month of sample collection (25% vs. 10%). Among the HIV-positive infants, a higher proportion at MZCH (92%) started antiretroviral therapy than at CDH (46%). The relative risk (RR) of death was higher among infants with late DBS collection (RR 1.3, 95%CI 1.0-1.7) or no collection (RR 5.8, 95%CI 4.6-7.2), and when guardians did not receive test results (RR 8.3, 95%CI 5.7-11.9). Conclusion: EID programmes performed equally poorly at both hospitals, and might be helped by point-of-care DNA PCR testing. Better programme implementation and active follow-up might improve infant outcome and retention in care.

Contexte: Hôpital central Mzuzu (MZCH), Mzuzu, et hôpital de district de Chitipa (CDH), Chitipa, Malawi.Objectif: Comparer la prise en charge et les résultats des nourrissons exposés au virus de l'immunodéficience humaine (VIH) dans les programmes de Diagnostic précoce du nourrisson (EID) au MZCH (test ADN réaction polymérase en chaîne [PCR] fait sur place) et au CDH (échantillons envoyés au MZCH) entre 2013 et 2014.Schéma: Etude rétrospective de cohorteRésultats: Parmi les nourrissons enrôlés au MZCH (n = 409) et au CDH (n = 176), les résultats d'ADN PCR ont été communiqués aux responsables des enfants dans 56% et 51% des cas, respectivement. Le délai médian du recueil de l'échantillon à la réception des résultats par les parents a été de 34 jours pour le MZCH et de 56 jours pour le CDH. Dans les deux hôpitaux, seulement la moitié des échantillons de sang séché (DBS) a été recueillie entre 6 et 8 semaines. Plus de parents du MZCH que du CDH ont reçu les résultats du test dans le mois suivant le recueil de l'échantillon (25% contre 10%). Parmi les nourrissons VIH positifs, une proportion plus élevée au MZCH (92%) a mis en route le traitement antirétroviral comparée au CDH (46%). Le risque relatif de décès a été plus élevé parmi les nourrissons ayant eu un recueil tardif de DBS (RR 1,3 ; IC95% 1,0­1,7) ou pas de recueil (RR 5,8 ; IC95% 4,6­7,2) et quand les parents n'ont pas reçu les résultats du test (RR 8,3 ; IC95% 5,7­11,9).Conclusion: Les programmes d'EID ont été aussi peu performants dans les deux hôpitaux et pourraient être améliorés par la possibilité de réaliser sur place le test PCR ADN. Une meilleure mise en œuvre du programme et un suivi actif pourraient améliorer les résultats pour les nourrissons et leur rétention en soins.

Marco de referencia: El Hospital Central de Mzuzu (MZCH) y el Hospital Distrital de Chitipa (CDH), en Malawi.Objetivo: Comparar el manejo y los desenlaces clínicos de los lactantes expuestos al virus de la inmunodeficiencia humana (VIH) en los programas de diagnóstico temprano del lactante (EID) en el MZCH (realización local de pruebas mediante la reacción en cadena de la polimerasa a partir de ADN [PCR-ADN]) y el CDH (muestras enviadas al MZCH) del 2013 al 2014.Método: Fue este un estudio retrospectivo de cohortes.Resultados: De los lactantes inscritos en el MZCH (n = 409), el resultado de la prueba PCR-ADN se comunicó a la persona encargada del niño en un 56% de los casos; esta proporción fue 51% en los lactantes inscritos en el CDH (n = 176). La mediana del lapso entre la obtención de la muestra y la entrega de los resultados a los encargados fue 34 días en el MZCH y 56 días en el CDH. En ambos hospitales, solo la mitad de las muestras de sangre seca (DBS) se recogió en 6 a 8 semanas. Más tutores de los lactantes en el MZCH que en el CDH recibieron el resultado de la prueba en el primer mes después de haber aportado la muestra (25% contra 10%). De los lactantes con resultado positivo frente al VIH, inició tratamiento antirretrovírico una mayor proporción de los niños atendidos en el MZCH (92%) que en el CDH (46%). El riesgo relativo (RR) de mortalidad fue más alto en los lactantes en quienes se obtuvo la muestra de DBS tardíamente (RR 1,3; IC95% de 1,0 a 1,7), en quienes no se obtuvo (RR 5,8; IC95% de 4,6 a 7,2) y cuando los tutores no recibieron los resultados (RR 8,3; IC95% de 5,7 a 11,9).Conclusión: El desempeño de los programas EID fue igualmente deficiente en ambos hospitales y se podría mejorar con la práctica de la prueba PCR-ADN en el momento de la atención. Una mejor ejecución del programa y un seguimiento activo contribuiría a obtener desenlaces clínicos más favorables en los lactantes y a retenerlos en los servicios de atención.

Expression of transforming growth factor-ß1 in neonatal rats with hyperoxia-induced bronchopulmonary dysplasia and its relationship with lung development.

The aim of this study was to detect the expression of transforming growth factor-ß1 (TGF-ß1) in neonatal rats with hyperoxia-induced bronchopulmonary dysplasia (BPD) and to explore its relationship with lung development. Forty-eight rats (2-3 days old) were randomly divided into a hyperoxia group and a control group (N = 24) which were then fed in ≥95% oxygen atmosphere and air, respectively. On the 1st, 3rd and 7th days of hyperoxia exposure, morphological changes of lung tissues were observed under an optical microscope. TGF-ß1 mRNA and protein levels in lung tissues were detected by real-time quantitative polymerase chain reaction and western blot, respectively. With increasing time of hyperoxia exposure, the hyperoxia group gradually suffered from pathological changes such as poor development of lung tissues, alveolar simplification, decrease in the number of alveoli, and hindered pulmonary microvascular development. On the 7th day of hyperoxia exposure, TGF-ß1 mRNA and protein levels (relative to b-actin) of the hyperoxia group (0.34 ± 0.19 and 0.21 ± 0.09, respectively) were significantly lower than those of the control group (0.83 ± 0.45 and 0.57 ± 0.45, respectively; P < 0.05). TGF-ß1 participates in the pathogenesis of BPD as an important regulatory factor during pulmonary vascular development.

Biomarkers of peritoneal fluid in endometriosis identified by surface-enhanced laser desorption/ionization time-of-flight.

OBJECTIVES: This work aims to detect the peritoneal fluid proteomic patterns in endometriosis patients, build diagnostic models, and evaluate its clinical significance. STUDY DESIGN: The authors used SELDI-TOF-MS protein chip array technology to detect biomarkers of peritoneal fluid in endometriosis patients. Fourteen endometriosis patients and 16 persons without endometriosis as control group were tested. RESULTS: Four potential biomarkers (4428m/z, 6891m/z, 13766m/z, and 6427m/z) were found. CONCLUSIONS: This method showed great potential in screening better biomarkers for endometriosis.

RNAi-mediated knockdown of FANCF suppresses cell proliferation, migration, invasion, and drug resistance potential of breast cancer cells

Fanconi anemia complementation group F protein (FANCF) is a key factor, which maintains the function of FA/BRCA, a DNA damage response pathway. However, the functional role of FANCF in breast cancer has not been elucidated. We performed a specific FANCF-shRNA knockdown of endogenous FANCF in vitro. Cell viability was measured with a CCK-8 assay. DNA damage was assessed with an alkaline comet assay. Apoptosis, cell cycle, and drug accumulation were measured by flow cytometry. The expression levels of protein were determined by Western blot using specific antibodies. Based on these results, we used cell migration and invasion assays to demonstrate a crucial role for FANCF in those processes. FANCF shRNA effectively inhibited expression of FANCF. We found that proliferation of FANCF knockdown breast cancer cells (MCF-7 and MDA-MB-435S) was significantly inhibited, with cell cycle arrest in the S phase, induction of apoptosis, and DNA fragmentation. Inhibition of FANCF also resulted in decreased cell migration and invasion. In addition, FANCF knockdown enhanced sensitivity to doxorubicin in breast cancer cells. These results suggest that FANCF may be a potential target for molecular, therapeutic intervention in breast cancer.

RNAi-mediated knockdown of FANCF suppresses cell proliferation, migration, invasion, and drug resistance potential of breast cancer cells.

Fanconi anemia complementation group F protein (FANCF) is a key factor, which maintains the function of FA/BRCA, a DNA damage response pathway. However, the functional role of FANCF in breast cancer has not been elucidated. We performed a specific FANCF-shRNA knockdown of endogenous FANCF in vitro. Cell viability was measured with a CCK-8 assay. DNA damage was assessed with an alkaline comet assay. Apoptosis, cell cycle, and drug accumulation were measured by flow cytometry. The expression levels of protein were determined by Western blot using specific antibodies. Based on these results, we used cell migration and invasion assays to demonstrate a crucial role for FANCF in those processes. FANCF shRNA effectively inhibited expression of FANCF. We found that proliferation of FANCF knockdown breast cancer cells (MCF-7 and MDA-MB-435S) was significantly inhibited, with cell cycle arrest in the S phase, induction of apoptosis, and DNA fragmentation. Inhibition of FANCF also resulted in decreased cell migration and invasion. In addition, FANCF knockdown enhanced sensitivity to doxorubicin in breast cancer cells. These results suggest that FANCF may be a potential target for molecular, therapeutic intervention in breast cancer.

The Bonebridge semi-implantable bone conduction hearing device: experience in an Asian patient.

For over three decades, bone conduction hearing aids have been changing the lives of patients with impaired hearing. The size, appearance and fitting discomfort of early generations of bone conduction hearing aids made them unpopular. The advent of bone-anchored hearing aids in the 1970s offered patients improved sound quality and fitting comfort, due to the application of osseointegration. However, the issue of post-operative peri-abutment pin tract wound infection persisted. The Bonebridge system incorporates the first active bone conduction device, and aims to resolve peri-abutment issues. Implantation of this system in an Asian patient is presented.

Microbiological and clinical characteristics of bacteraemia caused by the hypermucoviscosity phenotype of Klebsiella pneumoniae in Korea.

Hypermucoviscous (HV) isolates of Klebsiella pneumoniae have been linked to virulence potential in experimental infections. We examined 33 isolates of K. pneumoniae from patients with bacteraemia for the HV phenotype on agar culture, and determined their virulence potential by screening for capsular (K) serotype by polymerase chain reaction and the presence of seven virulence factor genes. Fourteen (42·4%) isolates expressed the HV phenotype and 11 of these were serotype K1 or K2; these serotypes were not identified in HV-negative isolates. The genes rmpA, rmpA2, aerobactin, wabG and allS were significantly more frequent in HV than non-HV isolates. Multilocus sequence typing identified 21 sequence types (ST), eight of which were found in HV-positive isolates and the clonal relatedness of isolates of the most frequent types (ST23 and ST11) from different hospitals was confirmed by pulsed-field gel electrophoresis. The HV phenotype was more associated with community-acquired infection with a lower frequency of fatal underlying illness, but with significantly more focal infections, notably liver abscesses. Clinicians should be aware of such clinical impacts of the HV phenotype.

Vibrant Soundbridge system: application of the stapes coupling technique.

INTRODUCTION: The use of the stapes coupling technique, employed in the Vibrant Soundbridge system, is technically less demanding than the vibroplasty technique, and is more likely to generate a positive outcome without significantly changing residual hearing or resulting in medical or surgical complication. METHOD: We report a patient with repeated left ossiculoplasty failure, who was successfully implanted with a Vibrant Soundbridge. CONCLUSION: We believe that the stapes coupling technique can provide natural stimulation to the inner ear, resulting in a better perceived sound quality.

Discovery of altered protein profiles in epithelial ovarian carcinogenesis by SELDI mass spectrometry.

OBJECTIVE: Identification of proteomic alterations in epithelial ovarian tumorigenesis may facilitate the understanding of progression of this disease. METHODS: Specific protein peak patterns were identified in 20 microdissected epithelial ovarian tumors (13 epithelial ovarian cancers (EOCs) and 7 low malignant potential (LMP) tumors), as well as in the matched normal cells. Protein profiles were generated by surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) from all the different types of cells. RESULTS: Among seven protein peaks from EOC cells, six were significantly increased while one was decreased compared with normal cells, and three peaks from LMP cells were markedly increased while one was decreased compared with normal cells. CONCLUSIONS: The combination of SELDI and laser capture microdissection (LCM) is effective in finding the key molecules in ovarian tumorigenesis. Further identification of these protein peaks is important and these malignant protein signatures lend themselves to identification of populations at high-risk for EOC and for monitoring response to EOC chemopreventive agents.