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Objective To explore effect of increasing cerebrospinal fluid (CSF) leptin level by administration from the foramen magnum to the cerebellum medulla oblongata pool in osteoporosis in rabbit model established by ovariectomy (OVX) and methylprednisolones.Methods According to certain inclusion and exclusion criteria,30 adult rabbits which were healthy 5-month-old female New Zealand white rabbits with the same batch and weight were divided into 5 groups (n=6) with random number table,including control group,normal saline group,Low dose group,middle dose group and high dose group.The animal model of osteoporosis (OP) was established 8 weeks after ovariectomy (bilateral ovarian resection) and intramuscular injection of methylprednisolone daily for 8 consecutive weeks in all groups.The control group were only simulated puncture and didn't inject any drugs or normal saline,the normal saline group were intracerebroventricular (ICV) injection with NS and Low,Middle and High dose group injected with low,middle and high dose of leptin respectively at 0,3,7 days after OVX.Before (0 week) and 4,8 weeks after the operation,the measure of dual energy X-ray absorptiometry was done on all the rabbits femur and bone mineral density (BMD) was delivered.Before (0 week) and 4,8 weeks after the first administration,leptin in cerebrospinal fluid (CSF) and serum growth hormone (GH),insulin growth factor 1 (IGF-1) and osteocalcin (OT/BGP) were tested with Enzyme linked immunosorbent assay and serum calcium,phosphorus and alkaline phosphatase (ALP) with automatic biochemical analyzer (ACA).The rabbits were sacrificed and the thalamus and femoral neck were harvested to test the expression of leptin receptor (LEPR) in thalamus and bone morphogenetic protein-2 (BMP-2) and type Ⅰ collagen (Col Ⅰ) at 8 weeks after the first administration.Results Compared with the control group and NS group,the cerebrospinal fluid (CSF) leptin concentration of leptin administration groups kept in a higher level significantly at 4 and 8 weeks after leptin administration,it was positively correlated with the concentration of administration.After administration 4 and 8 weeks,the bone mineral density (BMD) of leptin administration groups were not significantly lower than that of non-administration groups,and the high dose group was unchanged.The serum osteocalcin/bone glad protein (OT/BGP) concentration of leptin administration groups decreased more significantly compared with the non-administration groups at 4 weeks after leptin administration;and it rebounded more significantly than the non-administration groups and low dose group rebounded more significantly than other non-administration groups at 8 weeks.The concentration of serum growth hormone (GH) of 5 groups showed a downward trend at 4 and 8 weeks,the serum GH concentration of leptin administration groups decreased more significantly than the non-administration groups.The concentration of serum insulin-like growth IGF-1 of 5 groups were decreased significantly at 4 weeks,and the serum IGF-1 concentration of control and low dose groups were significantly higher than other 3 groups at 8weeks.Compared with the non-administration groups,the concentration of serum Calcium of leptin administration groups were increased more obvious,and it was positively correlated with the concentration of administration.Serum phosphorus of 5 groups were decreased at 4 weeks and elevated at 8 weeks;there was no difference among the groups.Serum ALP showed a continuous downward trend,and the middle dose group decreased significantly compared with other groups at 4 and 8 weeks.RTPCR results showed that the BMP-2 and Col Ⅰ mRNA expression increased and positively correlated with leptin administration concentration at 8weeks.Conclusion The increase of cerebrospinal fluid leptin level can promote the bone metabolism,reduce bone loss and precaution the occurrence of osteoporosis in rabbit model caused by OVX and methylprednisolone,and it was positively correlated with the concentration of administration.That provides a new method for the prevention and treatment of osteoporosis in rabbit model.
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Objective To determine the effect of induction of immune tolerance by intrathymic injection of myelin basic protein (MBP) for treatment of surgical brain injury (SBI) in rats.Methods Twenty-four male SD rats were divided into experimental group,control group and sham group with 8 rats each,according to the random number table.SBI model was established in experimental group and control group.Thymuses were exposed and injected with MBP (experimental group) or saline (control group) respectively.Sham group received craniotomy without durotomy and thymus paracentesis.Modified neurological severity score (MNSS),cerebral edema volume,levels of peripheral blood proinflammatory cytokine interleukin (IL)-2 and anti-inflammatory cytokine IL-4,and CD4 +/CD8 + T-cell ratio were measured at postoperative 1,3,7,14 and 21 d.Results MNSS between control and experimental groups differed significantly at each time point (P ＜ 0.05).Compared to sham group,MNSS in experimental group was recovered the same value at 21 d (P ＞ 0.05),but in control group remained high over the study (P ＜ 0.05).Less cerebral edema was observed in control and experimental groups than in control group at each time point (P ＜ 0.05),but all were lower than these in sham group (P ＜ 0.05).Lower IL-2 concentrations and higher IL-4 concentrations were observed in experimental group than in control group at 3,7 and 14 d (P ＜0.05).Compared to sham group,IL-2 concentrations in experimental group and IL-4 concentrations in control group revealed no significant differences at each time point (P ＞0.05).CD4 +/CD8 + T-cell ratio between experimental group and control group revealed significant difference at 1,7 and 14 d (P ＜0.05).CD4 +/CD8 + T-cell ratio in experimental group and control group recovered to the same level in sham group at 7 and 21 d(P ＞ 0.05).Conclusion Induction of immune tolerance through intrathymic injection of MBP can reduce neurological deficit and brain edema and facilitate the recovery of function from SBI.
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[ ABSTRACT] AIM:To investigate the effect of microRNA-375 ( miR-375) on the viability, cell cycle and apop-tosis of HCT116 cells.METHODS: The expression of miR-375 in different colorectal cancer cell lines was detected by real-time PCR.The miR-375 mimics was transfected into HCT116 cells by LipofectamineTM 2000.The mRNA expression of miR-375 and AEG-1 was detected by real-time PCR.The HCT116 cell viability was detected by MTT assay.The changes of apoptosis and cell cycle distribution were analyzed by flow cytometry.RESULTS:Real-time PCR showed that miR-375 expression was the lowest in HCT116 among 4 colorectal cancer cell lines.The expression level of miR-375 significantly in-creased in miR-375 mimics group compared with that in the negative control group.The high expression level of miR-375 significantly inhibited the mRNA expression of AEG-1.After transfection with miR-375 mimics, the cell viability was in-hibited, the apoptotic rate was increased, the proportion of G1-stage cells was increased, and the proportion of S-stage cells was decreased.CONCLUSION:miR-375 inhibits the viability, mediates the cell cycle arrest and promotes the apoptosis of colon cancer HCT116 cells.miR-375 may act as a tumor suppressor in colorectal cancer by inhibiting AEG-1.
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[ ABSTRACT] AIM:To investigate the characteristics of the intestinal microbial flora in the pregnant women with congenital heart disease fetus ( PW group) and normal pregnant women ( NW group) .METHODS: Stool samples were collected from 15 NW and 17 PW cases.The bacterial genomic DNA was extracted.The 16S rDNA was amplified by PCR, and the second generation of Illumina sequencing was conducted.RESULTS: We obtained 2 696 276 ( NW group) and 2 445 530 ( PW group) optimized sequences.The coverage was greater than 97%.We obtained 77 243 operational taxono-mic units ( OTUs) in NW group and 75 600 OTUs in PW group after a 97%similarity merge.In NW group, the Chao 1 in-dex and the Shannon index were greater than those in PW group.The diversity analysis of microbial population indicated that they were mainly composed of Firmicutes, Proteobacteria and Actinobacteria.In family, the Bifidobacteriaceae and Cori-obacteriaceae were significantly different through analysis of variance.CONCLUSION: The Bifidobacteriaceae and Cori-obacteriaceae may play an important role in the occurrence of congenital heart disease.
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Objective To investigate the enhanced osteogenesis during bone fracture healing after intracerebroven-tricular (ICV) leptin injection, using rabbit model with created segmental bone defect in right tibial. Method Segmental critical-sized defects were created at the right tibial bone of skeletally mature New Zealand white rabbits. In experiment group (leptin group), recombinant rabbit leptin was injected into cerebellomedullary cistern through foramen magnum. While in control group, normal saline was injected in the same way. Bone Mineral Density (BMD) was evaluated by qCT at 1st, 3rd, 7th, 14th and the 21st days. At the 21st days, all rabbits were euthanized to collect the right tibia for histomorphology, to ex-amine the BMP-2 expression in the bone callus by Fluorescence in situ hybridization (FISH). Result In the leptin group, body weight declined more obviously than control group then it start to arise at the 14th day;qCT showed significant higher BMD in leptin group than in control group at the 7th day;FISH showed a higher BMP-2 expression in leptin group than in control group. Conclusion Cerebellomedullary leptin injection through foramen magnum could accelerate limb fracture healing in rabbit model with right tibial bone defection.
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<p><b>BACKGROUND</b>Many researches demonstrate that the secondary brain injury which is caused by autoimmune attack toward brain antigens plays an important role in surgical brain injury (SBI). Although traditional immunosuppression can reduce autoimmune attack, it will lower the body immunity. Immune tolerance, by contrast, not only does not lower the body immunity, but also could lighten autoimmunity. This study used thymus tolerance to develop an immune system that is tolerant to autologous cerebrospinal fluid (CSF) and autologous brain tissue so that autoimmune injury can be suppressed following the disruption of the blood-brain barrier, thereby reducing brain damage.</p><p><b>METHODS</b>Eighty experimental rabbits were divided into five groups by random number table method: 16 in SBI group (group A), 16 in SBI+CSF drainage group (group B), 16 in SBI+CSF drainage+PBS injection group (group C), 16 in SBI+CSF drainage+CSF intrathymic injection group (group D), and 16 in SBI+brain homogenate intrathymic injection group (group E). Rabbits' CSF was drained in group B; was drained and injected PBS into thymus in group C; was drained and injected CSF into thymus in group D; and was injected brain homogenate in group E. Half of the rabbits in each group were phlebotomized on 1st, 3rd, 7th, and 14th days to observe the changes in IL-l, TGF-β by ELISA test, and CD4CD25 regulatory T cells ratio by flow cytometry, and in other animals brain tissues were taken on 7th day for exploring FasL expression by RT-PCR. The least significant difference (LSD) test was used to make paired comparisons; P < 0.05 was considered statistically significant.</p><p><b>RESULTS</b>The levels of FasL, TGF-β, and the ratios of CD4CD25 regulatory T cells in groups D and E were apparently higher than those in other three groups (P < 0.05). Likewise, the levels of IL-1 in these two groups were lower than the other three groups (P < 0.05). Moreover, the ratios of CD4CD25 regulatory T cells and the levels of TGF-β in groups B and C were higher than those in group A, but the level of IL-1 was lower than that in group A (P < 0.05). There was no significant difference between groups B and C, and groups D and E.</p><p><b>CONCLUSION</b>Thymic injection of CSF and brain homogenate may be able to reduce inflammation after SBI, so thymus immune tolerance may be a useful therapy to treat SBI.</p>
Subject(s)
Animals , Rabbits , Autoantigens , Brain , General Surgery , Brain Injuries , Therapeutics , Immune Tolerance , Physiology , Thymus Gland , Allergy and ImmunologyABSTRACT
The insecurity factors of third-class steel implants in the clinical application are introduced.A method for organizing the stocking way and improving the management in hospital are used to monitor and manage all the processes of the steel implants.All problems in management are solved,and this can insure the operation security and depress the costs of patients.