Animals ; Benzodiazepines ; pharmacology ; Ducks ; physiology ; Growth Hormone ; blood ; Insulin-Like Growth Factor I ; metabolism ; Serum ; metabolism ; Weight Gain ; drug effects

BACKGROUND:Semaphorin7A (Sema7A) is a kind of cel surface protein, which can promote the fusion of osteoclasts and the migration of osteoblasts at the same time, affecting the dynamic balance of the bone. It is speculated that Sema7A siRNA may inhibit osteoblast apoptosis induced by titanium particles. OBJECTIVE:To study the effect of Sema7A on the preosteoblast activity inhibited by titanium particles. METHODS:Mouse MC3T3-E1 preosteoblasts at passages 6 and 7 were divided into four groups: in blank control group, MC3T3-E1 cels were cultured alone; in standard control group, cel were cultured with titanium particles; in experimental groups 1 and 2, the cels were cultured with titanium particles+Sema7A overexpression plasmids and titanium particles+Sema7A siRNA, respectively. Apoptotic rate of MC3T3-E1 cels was detected by flow cytometry; the mRNA expression of bone sialoprotein, osteocalcin and type I colagen was detected by Q-PCR; western blot assay was adopted to detect the protein expression of bone sialoprotein, osteocalcin and type I colagen; alizarin red calcium nodule staining was taken to detect the degree of osteoblast mineralization. RESULTS AND CONCLUSION:The expressions of bone sialoprotein, osteocalcin and type I colagen were decreased in the standard control group and experimental group 1, but these expression were significantly increased in the experimental group 2 compared with the standard control group (P < 0.05). Flow cytometry results suggested that the apoptotic rate of osteoblasts in the experimental group 1 was significantly higher than that in the other groups (P < 0.05), and the apoptotic rate in the experimental group 2 was lower than that in the standard control group (P < 0.05). Alizarin red staining showed that there were no obvious mineralized nodules in the experimental group 1, but mineralized nodules formed in the experimental group 2. In brief, the genetic interference technique that inhibits the activity of Sema7A can interfere the process of mouse MC3T3-E1 preosteoblast differentiation inhibited by titanium particles, and thus provide a feasible way for the clinical treatment of wear particles-induced osteolysis using biotechnology.

BACKGROUND:Sema7A is a kind of cel surface protein, which can promote the fusion of osteoclasts and the migration of osteoblast at the same time, affecting the dynamic balance of bone. OBJECTIVE:To investigate whether Sema7A siRNA has ainhibitory effect on the osteoclast activation in the process of osteolysis which induced by titanium particles. METHODS:The precursor osteoclasts with the concentration of 4×109 RESULTS AND CONCLUSION:At 7 days of culture, the expression levels of interleukin-1, interleukin-1β, tumor necrosis factor α, matrix metaloproteinase-9 and the receptor activator of nuclear factor-κB in the positive control, /L were seeded on 96-wel plates containing glass cover slips, and divided into four groups: blank control, positive control, experiment and negative control groups. The cel culture medium was added into the control group. 20 μL un-transfected siRNA supernatant was added into the positive control group. 20 μL transfected Sema7A siRNA supernatant was added into the experiment group. 20 μL transfected control siRNA supernatant was added into the negative control group. The supernatant was obtained through the co-culture between titanium particles solution and monocyte-macrophage cel line RAW264.7of mouse for 24 hours. siRNA was transfected into mononuclear macrophage cel lines RAW264.7 of mice. negative control and experiment groups were higher than those in the control group (P < 0.05). The expression level of each factor in the experiment group was lower than that in the positive control and negative control groups (P < 0.05). At 8 days of culture, the proliferation activity of osteoclasts and the number of positive cels stained by tartrate-resistant acid phosphatase in the positive control, negative control and experiment groups were higher than those in the control group (P < 0.05). The proliferation activity of osteoclasts and the number of positive cels stained by tartrate-resistant acid phosphatase in the experiment group were lower than those in the control and negative groups (P < 0.05). These results demonstrate that Sema7A siRNA has a certain inhibitory effect on the osteoclast activation induced by titanium particles.

Carcinoma, Medullary ; complications ; pathology ; Humans ; Male ; Middle Aged ; Thyroid Neoplasms ; complications ; pathology

BACKGROUND:Previous studies have indicated that ulinastatin can inhibit RANKL-induced osteoclastogenesis on RAW264.7 cells and also lower matrix metal oproteinase-9 expression and activity. However, it remains be unclear whether ulinastatin has the intervention effect on polymethyl methacrylate (PMMA)-induced MC3T3-E1 mouse preosteoblast apoptosis or not. ＜br> OBJECTIVE:To explore the intervention role of ulinastatin on the PMMA-induced MC3T3-E1 mouse preosteoblast apoptosis and its effects on type I col agen, osteocalcin, matrix metal oproteinase-2 mRNA expression. ＜br> METHODS:MC3T3-E1 mouse preosteoblasts at passages 6 and 7 were divided into four groups:blank group (only cultured MC3T3-E1 mouse preosteoblast), PMMA-induced group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension), low dose ulinastatin group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension+500 U/mL ulinastatin) and high dose ulinastatin group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension+5 000 U/mL ulinastatin). MTT method was adopted to detect the proliferation activity of proliferative activity of MC3T3-E1 mouse preosteoblast;alizarin red staining method was used to observe mineralization nodules of MC3T3-E1 mouse preosteoblast among different groups;the change of apoptosis rate for MC3T3-E1 cells was detected by flow cytometry analysis;semi-quantitative RT-PCR was taken to analyze type I col agen, osteocalcin, matrix metal oproteinase-2 mRNA expression level in MC3T3-E1 mouse preosteoblasts among different groups. ＜br> RESULTS AND CONCLUSION:Compared with the blank group, PMMA significantly inhibited the proliferation activity of MC3T3-E1 mouse preosteoblast (P＜0.05), and however significantly promoted cells apoptosis (P＜0.05). After addition of different concentrations of ulinastatin (500, 5 000 U/mL), the proliferation activity of MC3T3-E1 mouse preosteoblasts significantly raised (P＜0.05), and cells apoptosis rate significantly decreased (P＜0.05), showing the dose and time-dependent relation. Type I col agen and osteocalcin mRNA expression levels both significantly decreased after co-culture in PMMA group compared with the blank group (P＜0.05), matrix metal oproteinase-2 mRNA expression level, however, significantly increased (P＜0.05). After intervention with 5000 U/mL ulinastatin, type I col agen and osteocalcin mRNA expression levels both significantly increased, while matrix metal oproteinase-2 mRNA expression level significantly decreased (P＜0.05). PMMA group showed no obvious mineralization nodules. Yet, mineralization nodules were formed in the blank group, high and low dose ulinastatin groups. These results indicate that ulinastatin could have the inhibitory effect on the PMMA-induced MC3T3-E1 mouse preosteoblast apoptosis, and it could promote type I col agen and osteocalcin mRNA expression and yet suppress matrix metal oproteinase-2 mRNA expression.

Antigens, CD20 ; analysis ; Bone Marrow Transplantation ; adverse effects ; CD79 Antigens ; analysis ; Female ; Humans ; Immunohistochemistry ; Liver Transplantation ; adverse effects ; Lymphoma, Large B-Cell, Diffuse ; etiology ; metabolism ; pathology ; Lymphoproliferative Disorders ; etiology ; metabolism ; pathology ; Male ; Middle Aged ; Postoperative Complications ; etiology ; metabolism ; pathology ; Young Adult

BACKGROUND:It is presumed that urinary trypsin inhibitor could have protective effects on local and systemic tissues and could inhibit osteoclast proliferation and activation under long-term chronic inflammation conditions and in ischemic and anoxic environment which was induced by prosthetic wear. OBJECTIVE:To investigate the inhibitory effect of ulinastatin on receptor activator for nuclear factor-κb ligand-induced differentiation, proliferation and osteoclastogenesis of RAW264.7 cells and its effects on matrix metal oproteinase-2, matrix metal oproteinase-9 expression level and activity. METHODS:Mouse monocyte/macrophage cellline RAW264.7 was treated with different concentrations of urinary trypsin inhibitor (0, 500, 5 000 U/mL) for 24, 48 and 72 hours. Experiments were divided into four groups:the blank group (RAW264.7 cells), receptor activator for nuclear factor-κb ligand-induced group (0 U/mL ulinastatin), 500 U/mL ulinastatin group and 5 000 U/mL ulinastatin group. RESULTS AND CONCLUSION:(1) MTT results indicated that there was no significant difference on the proliferation of RAW264.7 cells treated with urinary trypsin inhibitor at 0-5 000 U/mL (P>0.05) (2) Tartrate-resistant acid phosphatase staining results revealed that compared with receptor activator for nuclear factor-κb ligand-induced group, the number of tartrate-resistant acid phosphatase-positive cells was significantly less in the ulinastatin group (P<0.05), showing a time-dose dependent manner. (3) Immunohistochemisical results found that compared with receptor activator for nuclear factor-κb ligand-induced group, the percentage of matrix metal oproteinase-9-positive cells was apparently lower in the ulinastatin group. (4) Western blot assay results demonstrated that matrix metal oproteinase-9 expression was low in the RAW264.7 cells alone. At 48 hours after addition of receptor activator for nuclear factor-κb ligand, matrix metal oproteinase-9 protein expression was large. At 72 hours after culture in the 5 000 U/mL ulinastatin group, matrix metal oproteinase-9 protein expression was evidently reduced. (5) Gelatin zymography results showed that compared with the receptor activator for nuclear factor-κb ligand-induced group, matrix metal oproteinase-9 expression was significantly lower in the 5 000 U/mL ulinastatin group (P<0.05). Results suggested that urinary trypsin inhibitor inhibited receptor activator for nuclear factor-κb ligand-induced osteoclastogenesis and diminished matrix metal oproteinase-9 expression and activity.

Objective To investigate the clinical effect of type A botulinum toxin and nipple retractor in correcting severe inverted nipples.Methods All the patients with bilateral severe inverted nipples that sought consulting in the Plastic and Reconstructive Surgery Center of Peking Union Medical College Hospital were included in this study and the patients were randomized into different treatment group with nipple retractor only or BTX-A (Lanzhou,China) combined with nipple retractor.In the combined therapy group,50 u BTX-A was injected into the bottom of each nipple.2 weeks later,nipple retractor was placed and kept wearing for 6 months.For the retractor only group,no BTX-A was used.All the patients were followed up at 1 month,3 months and 6 months after the operation.Nipple projection was measured according to the profile view of pre-and post-operation.Effectiveness and complication rate were compared between the two groups.Results 20 patients were included in this study.Average nipple projection in the combined therapy group was (1.12±0.13) cm,(0.95± 0.10) cm and (0.73±0.11) cm (3 months,6 months and 12 months post-operation,respectively),which had a significant difference from that of the retractor group [(0.81±0.10) cm,(0.72±0.12) cm and (0.53±0.10) cm].Total complication rate of combined group was also lower than that of retractor group.The complications of the combined therapy group included hypopigmentation (1 case),without skin ulcer or wire dislocation.However in the retractor group,complications included skin ulcer (2 cases),hypopigmentation (1 case) and dislocation (1 case).No severe complications such as nipple necrosis happened in both groups.Conclusions BTX-A combined with nipple retractor is an effective method in correcting severe inverted nipple with low complication rate.

ObjectiveTo evaluate the diagnostic performance of CTCA in predicting myocardial perfusion defects through comparative analysis between MPI defects and severity of coronary stenosis on CTCA.MethodsFour hundred and seventy-eight patients who underwent CTCA and 99Tcm-MIBI MPI in the same period were analyzed retrospectively.According to the severity of coronary stenosis judged by visual evaluation of the vessel diameter,the patients were divided into five groups:no stenosis,mild stenosis,moderate stenosis,severe stenosis and total occlusion.MPI results were classified as negative or positive for perfusion defects,and the prevalence of perfusion defects in every group was calculated per-patient and per-vessel basis.The cut-off of stenotic severity for predicting myocardial perfusion defects was designated as 50％ or 75％,with MPI as standard reference.True positive,true negative,false positive and false negative statistics were thus determined separately on patient and vessel basis.The diagnostic performance for CTCA were calculated and compared.Pearson Chi-square and its partition tests or Fisher exact test were used to compare ordinal variables.ResultsFifty-eight patients showed myocardial perfusion defects.Either by patientbased or vessel-based analysis,the prevalence of myocardial perfusion defects showed an increasing trend with greater coronary artery stenosis in each group,and there were statistical differences among them (x2 =116.62 and 483.83,both P ＜ 0.05).On patient-based analysis,sensitivity ( SN),specificity ( SP),positive predictive value( PPV),negative predictive value(NPV) and accuracy (AC) for CTCA predicting myocardial perfusion defects were 62.1 ％ ( 36/58 ) and 34.5％ ( 20/58 ) (x2 =8.84,P ＜ 0.05 ),84.5％(355/420) and 97.1％ (408/420) (x2 =40.16,P ＜0.05),35.6％ (30/101) and 62.5％ (20/32) (x2 =7.19,P＜0.05),94.2％ (355/377) and 91.5％ (408/446) (x2 =2.18,P ＞0.05),81.8％ (391/478)and 89.5 ％ (428/478) (x2 =11.66,P ＜ 0.05 ) when the cutoff was set to 50％ and 75％,respectively.On vessel-based analysis,the SN,SP,PPV,NPV and AC for CTCA predicting myocardial perfusion defects were 58.8％ (40/68) and 30.9％ (21/68) (x2 =10.73,P ＜ 0.05),95.9％ (1768/1844) and 99.0％ (1826/1844) (x2 =36.72,P ＜ 0.05 ),34.5％ (40/116) and 53.8％ (21/39) (x2 =4.59,P ＜0.05 ),98.4％ (1768/1796) and 97.5％ ( 1826/1873 ) (x2 =4.14,P ＜ 0.05 ),94.6％ ( 1808/1912 ) and 96.6％ ( 1847/1912 ) (x2 =10.31,P ＜ 0.05 ),respectively.ConclusionsThe prevalence of myocardial perfusion defects correlates positively with the severity of coronary stenosis seen on CTCA.CTCA may predict perfusion defects with high SP and NPV.However,the PPV of CTCA in predicting myocardial perfusion defects is poor when the stenosis cut-off is set at 50％.It is significantly improved when the cutoff value is set at 75 ％.

OBJECTIVE: To study the percentages of polymorphonuclear leukocytes (PMN), mononuclear cells (MNC) and fibroblastic cells (FBC) in different post-traumatic intervals after skeletal muscle mechanical injury in rats. METHODS: The rat model of skeletal muscle mechanical injury was established. The rats were divided into injured groups (6 h, 12 h, 1 d, 3 d, 7 d, 10 d and 14 d after injury) and control group. The percentages of PMN, MNC and FBC in different post-traumatic intervals after skeletal muscle mechanical injury were assessed with HE staining and image analysis. RESULTS: At post-injury 6-12h, the percentages of PMN and MNC infiltration appeared in injured sites and that of PMN reached peak. At 1 d, the percentage of MNC infiltration appeared and reached peak, while that of PMN decreased. At 3-7 d, the percentage of FBC gradually increased, while that of PMN and MNC decreased. At 10-14d, the percentage of FBC reached peak. CONCLUSION The percentages of PMN, MNC and FBC in injured zones showed time-dependent changes, which might be used as reference index for determination of age of skeletal muscle injury.
Animals ; Fibroblasts ; Muscle, Skeletal/injuries* ; Neutrophils ; Rats ; Time Factors