Longitudinal skeletal growth and growth plate morphological characteristics of chondro-tissue specific CUL7 knockout mice.

BACKGROUND: 3 M syndrome is first reported in 1975，which characterized by severe pre- and postnatal growth retardation, skeletal malformation and facial dysmorphism. These three genes (CUL7, OBSL1 and CCDC8) have been identified to be respond for 3 M syndrome, of which CUL7 is accounting for approximately 70%. To date, the molecular mechanism underlying the pathogenesis of 3 M syndrome remains poorly understood. Previous studies showed that no Cul7-/- mice could survive after birth, because of growth retardation at late gestational stage and respiratory distress after birth. The establishment of the animal model of cartilage specific Cul7 knockout mice (Cul7fl/fl;Col2a1-CreERT2 mice) has confirmed that Cul7fl/fl;Col2a1-CreERT2 mice can be selective in a time- and tissue-dependent manner, which can provide an experimental basis for further research on severe genetic diseases related to growth plates. OBJECTIVE: To establish a model of Cul7fl/fl;Col2a1-CreERT2 mice based on Cre/LoxP system, and to further observe its phenotype and morphological changes in growth plate. METHODS: The Cul7fl/fl;Col2a1-CreERT2 mice were taken as the experimental group, while the genotype of Cul7fl/+;Col2a1-CreERT2 mice were used as the control group. The gross morphological features and X-ray films of limbs in the two groups were observed every week for 3-6 consecutive weeks, and the length of the mice from nose to the tail, the length of femur and tibia were recorded. In the meantime, The histological morphology of tibial growth plates was compared between the two groups. RESULTS: A preliminary model of Cul7fl/fl;Col2a1-CreERT2 mice was established. The Cul7fl/fl;Col2a1-CreERT2 mice had abnormally short and deformed limbs (P<0.05), increased thickness of growth plate, the disorderly arranged chondrocyte columns, decreased number of cells in the proliferation zone, changes in the shape from flat to round, obviously expanded extracellular matrix, and disordered arrangement, thickening and loosening of bone trabecula at the proximal metaphysis of the femur. CONCLUSIONS: The knockout of Cul7 gene may affect both the proliferation of chondrocytes and the endochondral osteogenesis, confirming that Cul7 is essential for the normal development of bone in the body.

A Reliable Murine Model of Disseminated Infection Induced by Talaromyces Marneffei.

Talaromycosis (penicilliosis) caused by Talaromyces marneffei is one of the most important opportunistic infection diseases in tropical countries of South and Southeast Asia. Most infections occurred in individuals with human immunodeficiency virus (HIV) and the primarily reason for the increase in the number of the cases is HIV pandemic. The pathogenesis of T. marneffei infection is unclear. There is still no ideal animal model for studying talaromycosis. In this study, we developed a stable, safe and maneuverable murine model that mimics human T. marneffei disseminated infection using T. marneffei yeast intraperitoneal injected to BALB/c nude mice. We successfully observed symptoms similar to those seen in clinical patients in this murine model, including skin lesions, hepatosplenomegaly, pulmonary infection and mesenteric lesions. We further studied the pathological changes of various tissues and organs in the infected animals to help better understand the severity of the infection. This model may provide a good tool for studying disseminated infection induced by T. marneffei.

Post-menopausal Osteoporosis and Probiotics.

Postmenopausal osteoporosis (PMO) is characterized by low bone mass and structural deterioration of bone tissue with increased risk of fracture in postmenopausal women. It is due to the deficiency of estrogen production after menopause, which causes the imbalance in the bone remodeling process where resorption/formation skewed more towards resoption, which leads to bone loss. It causes high morbidity and severe health complication among the affected women. The current PMO therapy has many unwanted side effects and even increases the possibility of tumorigenesis. Therefore, an alternative therapy that is safe and effective is required. Probiotics are dietary supplements consisting of beneficial microbes and when administered in an adequate amount, confer a health benefit to the host. Recent scientific evidences suggested the link between the intestinal microbiota and bone health. This review discusses the process of bone remodeling and the role of intestinal microbiota on the bone metabolism of the host. Further, it summarizes the recent studies of probiotic on an animal model of PMO and also in post postmenopausal women.

Experimental Infection of Mid-Gestation Pregnant Female and Intact Male Sheep with Zika Virus.

Zika virus (ZIKV) is an arbovirus that causes birth defects, persistent male infection, and sexual transmission in humans. The purpose of this study was to continue the development of an ovine ZIKV infection model; thus, two experiments were undertaken. In the first experiment, we built on previous pregnant sheep experiments by developing a mid-gestation model of ZIKV infection. Four pregnant sheep were challenged with ZIKV at 57-64 days gestation; two animals served as controls. After 13-15 days (corresponding with 70-79 days of gestation), one control and two infected animals were euthanized; the remaining animals were euthanized at 20-22 days post-infection (corresponding with 77-86 days of gestation). In the second experiment, six sexually mature, intact, male sheep were challenged with ZIKV and two animals served as controls. Infected animals were serially euthanized on days 2-6 and day 9 post-infection with the goal of isolating ZIKV from the male reproductive tract. In the mid-gestation study, virus was detected in maternal placenta and spleen, and in fetal organs, including the brains, spleens/liver, and umbilicus of infected fetuses. Fetuses from infected animals had visibly misshapen heads and morphometrics revealed significantly smaller head sizes in infected fetuses when compared to controls. Placental pathology was evident in infected dams. In the male experiment, ZIKV was detected in the spleen, liver, testes/epididymides, and accessory sex glands of infected animals. Results from both experiments indicate that mid-gestation ewes can be infected with ZIKV with subsequent disruption of fetal development and that intact male sheep are susceptible to ZIKV infection and viral dissemination and replication occurs in highly vascular tissues (including those of the male reproductive tract).

Establishment of xenografts of urological cancers on chicken chorioallantoic membrane (CAM) to study metastasis.

Cancer of the urological system commonly occurs in the kidney, bladder, and prostate gland. The clear cell subtype of renal cell carcinoma (ccRCC) constitutes the great majority of kidney cancer. Metastatic ccRCC portends a very poor outcome with no effective treatment available. Prostate cancer is the most common cancer in males in the US. Despite recent advances in selective kinase inhibitors and immunotherapies, the rate of developing new treatment from bench to bedside is slow. A time-consuming step is at the animal drug testing stage, in which the mouse model is the gold standard. In the pursuit to streamline the in vivo cancer biology research and drug development, we explored the feasibility of the chicken chorioallantoic membrane (CAM) model to establish xenografts. The CAM model greatly shortens the time of tumor growth and lowers the cost comparing to immunocompromised mice. We generated CAM xenografts from ccRCC, bladder and prostate cancer, with established cancer cell lines and freshly isolated patient-derived tissues, either as primary tumor cells or small pieces of tumors. The successful CAM engraftment rate from the different tumor sources is 70% or above. Using our previously established metastatic ccRCC mouse model, we showed that the CAM xenograft maintains the same tumor growth pattern and metastatic behavior as observed in mice. Taken together, CAM can serve as a valuable platform to establish new patient-derived xenografts (PDXs) to study tumor biology, thus accelerating the development of individualized treatment to halt the deadly metastatic stage of cancer.

Parietal peritoneum graft for duodenum injuries in an animal model / Enxerto de peritônio parietal em lesão duodenal: modelo animal

ABSTRACT Background: Duodenal injuries and their surgical procedure cause a high morbidity and mortality. Aim: To assess the overall effectiveness of the auto-graft of peritoneum in the treatment of the perforation of the duodenum, aiming to reduce surgery time, costs, complexity and mortality. Methods: Twelve New Zealand rabbits, ages 4-6 months, both sexes, underwent designed surgical grade III duodenal injuries that were repaired 18 h after. Rabbits were surgically treated with the proposed auto-graft of peritoneum. Results: No postoperative deaths were observed; the animals presented corporal weight increase and were euthanized six months later. There was no significant difference between both groups relating to the postoperative evolution or in the histological changes. Conclusion: Auto-graft of the peritoneum and posterior fascia is a useful option for duodenal repair and that is worth of evaluation for humans.

RESUMO Racional: Lesões duodenais e seu procedimento cirúrgico causam alta morbimortalidade. Objetivo: Avaliar a eficácia geral de retalho peritoneal no tratamento da perfuração do duodeno, visando reduzir o tempo, os custos, a complexidade e a mortalidade cirúrgicas. Métodos: Doze coelhos da raça Nova Zelândia, com idades entre 4-6 meses, ambos os sexos, foram submetidos a lesões duodenais cirúrgicas de grau III, que foram reparadas 18 h depois. Coelhos foram tratados cirurgicamente com a proposta de auto-enxerto de peritônio. Resultados: Não foram observados óbitos pós-operatórios; os animais apresentaram aumento de peso corporal e foram eutanasiados seis meses depois. Não houve diferença significativa entre os dois grupos em relação à evolução pós-operatória ou nas alterações histológicas. Conclusão: A auto-enxertia do peritônio e da fáscia posterior é uma opção útil para o reparo duodenal e vale a pena ser avaliada em seres humanos.

Comprehensive training of rat abdominal aorta and common iliac artery for neurosurgical microvascular anastomosis / 中国脑血管病杂志

Objective To establish an animal model suitable for neurosurgeons for the comprehensive training of microvascular anastomosis using rat abdominal aorta and common iliac artery. Methods Twelve adult SD rats were selected,they were generally anesthetized and laparotomized.The abdominal aortas and bilateral common iliac arteries were exposed and fully separated.The lengths and diameters of abdominal aortas and common iliac arteries of each segment were measured.The end-to-end anastomosis were performed choosing the main trunk of the abdominal aorta without a branching artery and longer segment.The unilateral common iliac artery and the sacral median artery were used for end-to-side anastomosis.The bilateral common iliac arteries were used for end-to-side and side-to-side anastomosis.The micro Doppler ultrasound probe was used to detect the blood flow patency of each anastomotic stoma. Results Anatomical separation of the abdominal aortas and the common iliac arteries was successfully performed in 12 SD rats.Each rat could provide 4 vascular anastomosis exercises.The length of abdomen aorta trunk was 15.6 ± 2.5 mm and the diameter was 1.6 ±0.2 mm between the lower left renal artery and right iliolumbar artery.The side-to-side anastomosis was performed.The mean diameter of the median sacral arteries was 0.80 ±0.08 mm.After the fish mouth-like cutting,the end-to-side anastomosis of the right common iliac arteries were conducted.The lenth of left common iliac artery was 9.0 ± 1.5 mm,the diameter was 1.0 ± 0.1 mm,and was cut off at its origin and then the end-to-side anastomosis of the right common iliac arteries were conducted.After end-to-side anastomosis of bilateral common iliac arteries,its distal segment was arranged in parallel with a length of 5.1 ± 0.3 mm,and then the side-to-side anastomosis could be conducted. Conclusions The rat abdominal aorta and iliac artery model can be comprehensively used to simulate the commonly used neurosurgery bypass graft.It is suitable for neurosurgeons with a certain microsurgical basis to conduct a preliminary vascular anastomosis training.

Separation anxiety: at the neurobiological crossroads of adaptation and illness.

Physiological and adaptive separation anxiety (SA) is intimately connected with the evolutionary emergence of new brain structures specific of paleomammalians, the growth of neomammalian--and later hominid--brain and skull size, and the appearance of bipedalism. All these evolutionary milestones have contributed to expanding the behavioral repertoire and plasticity of prehuman and human beings, at the cost of more prolonged dependency of the infant and of the child on parental care. Separation anxiety disorder (SAD) can be seen as an exaggerated/inappropriate manifestation of SA that constitutes a gateway to poorer mental and physical health. By blending epidemiological, genetic-epidemiological, endophenotypic, and animal laboratory approaches, it is possible to delineate some of the mechanisms that link childhood-adolescence SA and SAD to health problems later in life. Causal mechanisms include gene-environment interplays and likely differential regulation of genes and functional net-works that simultaneously affect multiple behavioral and physical phenotypes after exposure to early-life adversity, including parental separation/loss.

La ansiedad de separación (AS) fisiológica y adaptativa está íntimamente relacionada con el surgimiento durante la evolución de nuevas estructuras cerebrales específicas de los paleomamíferos, el aumento del tamaño del cerebro y del cráneo de los neomamíferos y más tarde de los homínidos, y la aparición de la bipedestación. Todos estos hitos evolucionistas han contribuido a la expansión del repertorio conductual y a la plasticidad de los seres prehumanos y humanos, a costa de una dependencia más prolongada del lactante y del niño respecto al cuidado parental. El trastorno por ansiedad de separación (TAS) se puede considerar como una manifestación exagerada/inapropiada de la AS que constituye una puerta de entrada a una peor salud mental y física. Al combinar aproximaciones epidemiológicas, genético-epidemiológicas, endofenotípicas y de animales de laboratorio es posible bosquejar algunos de los mecanismos que relacionan la AS de la niñez-adolescencia y el TAS con problemas de salud en años posteriores. Los mecanismos causales incluyen interjuegos entre genes y ambiente y una probable regulación diferencial de genes y redes funcionales que afectan simultáneamente múltiples fenotipos conductuales y físicos después de la exposición a la adversidad temprana en la vida, incluyendo la separación y la pérdida parental.

L'anxiété de séparation (AS) physiologique et adaptée est intimement liée à l'émergence évolutive des nouvelles structures cérébrales spécifiques des paléomammifères, à la croissance du cerveau néomammifère, et plus tard hominidé, à la taille du crâne et à l'apparition de la bipédie. Tous ces jalons évolutifs ont contribué à élargir la plasticité et le répertoire comportementaux des êtres préhumains et humains, au prix d'une dépendance plus longue du nourrisson et de l'enfant au soin parental. Le trouble anxiété de séparation (TAS) peut être considéré comme une manifestation exagérée/inappropriée de l'AS, constituant une passerelle vers une dégradation de la santé mentale et physique. En associant les approches épidémiologiques, génético-épidémiologiques, endophénotypiques et des laboratoires animaux, il est possible de définir certains des mécanismes qui relient l'AS de l'enfance et de l'adolescence et les TAS aux problèmes de santé survenant plus tard dans la vie. Des interactions gène-environnement et probablement une régulation différentielle des gènes et des réseaux fonctionnels font partie du mécanisme causal, touchant simultanément de nombreux phénotypes comportementaux et physiques après une exposition à l'adversité dans la petite enfance, comme la séparation/perte d'un parent.

The application of modified shortened scope for ERCP procedure in swine animal model / 中华消化内镜杂志

Objective To evaluate modified shortened scope for ERCP procedure in swine animal model.Methods A total of 17 male pigs were divided into the conventional endoscope group (shortened scope used on human,n =8) and the modified endoscope group (modified shortened scope,n =9).The success rates,time of access to the pylorus,biliary or pancreatic orifice,the cannulation time of biliary or pancreatic duct,the stenting time of biliary or pancreatic duct,the total time of procedure and the X-ray exposure time were compared between the two groups.Results The success rates were 2/8 and 9/9 in the conventional endoscope group and the modified endoscope group,respectively(P =0.002).There were significantly differences of the cannulation time of biliary duct(180.00 ± 37.89 s VS 55.22 ± 11.56 s,P =0.000),cannulation time of pancreatic duct(55.22 ± 11.56 s VS 56.56 ±29.19 s,P =0.015),the time of access to pancreatic orifice(21.50 ±7.33 min VS4.67 ±2.63 min,P =0.000) and the total time of procedure(55.13 ±5.64 min VS 31.67 ±8.25 min,P =0.000) between the two groups.There were no significant differences in the time of passing through the pylorus,the time of finding biliary orifice,the stenting time of biliary or pancreatic duct or the X-ray exposure time between the two groups.Conclusion The modified shortened scope can improve the success rate of short scope position,facilitate locating biliary or pancreatic orifice,improve the success rate of pancreatic duct cannulation and shorten the cannulation time of biliary or pancreatic duct and the total operation time.

Ultrasound monitoring in the establishment of the model of alcoholic fatty liver in rabbits / 医学研究生学报

Objective The incidence of alcoholic fatty liver increases year by year in recent years .The aim of this study was to establish an animal model of AFL to investigate the pathogenesis of hepatic fibrosis . Methods This study involved 40 male Japa-nese rabbits aged (17.01 ±1.54) d and weighing 1.00-1.52 kg, which were equally randomized to an experimental group and a control group.The animals in the former group received lavage of 10 mL of 50%ethanol twice a day, with normal feedstuff and water, while those in the control group received normal feedstuff and water only .We performed ultrasonography for dynamic liver presentation before and at 12, 16, and 20 weeks after feeding, followed by pathological observation of the livers . Results After 12 weeks of eth-anol garage , fatty liver was observed in 18 of the rabbits and it deteriorated with the prolonged time of administration . The body weight was significantly decreased in the experimental rabbits as com-pared with the controls at 16 weeks ([2.48 ±0.30] vs [2.78 ± 0.15] kg, P vealed similar results (1 mild, 4 moderate, and 12 severe cases of fatty liver) in the former group.At 20 weeks, alcoholic fatty liver was found mainly in the S3-S4 stage. Conclusion Alcoholic fatty liver models could be successfully established in rabbits by etha-nol garage and ultrasonography is useful for monitoring the development and progression of the condition .