BACKGROUND:Osteoprotegerin and nitrogen monoxidum play a key role in the prevention and treatment of osteoporosis. However, the correlation of the two in inhibiting the proliferation and differentiation of osteoclasts remains unclear.
OBJECTIVE:To observe the effect of different doses of osteoprotegerin on nitric oxide production and endothelial nitric oxide synthase activity in osteoclasts.
METHODS:Tartrate resistant acid phophatase staining was used to test whether the induced cells are osteoclasts. Osteoclasts were divided into six groups:blank control group (no reagent);negative control group (DMEM);four osteoprotegerin groups (10, 25, 50, 75μg/L of osteoprotegerin). Annexinv-FITC kit and flow cytometry were used to test the apoptosis rate of osteoclasts. Real-time PCR was used to detect the expression of osteoclasts marker gene, TRAP mRNA and protein kinase K mRNA. Nitric oxide production and nitric oxide synthase activity were determined using the corresponding kits. Four osteoprotegerin groups were added with L-NAME, a kind of nitric oxide synthase inhibitor, to test the changes of the apoptosis rates of osteoclasts and the changes of the expression of TRAP mRNA and protein kinase K mRNA of osteoclasts.
RESULTS AND CONCLUSION:Osteoprotegerin inhibited the differentiation of osteoclasts and induced the apoptosis. Osteoprotegerin concentration had a positive correlation with the apoptosis rate of osteoclasts, and a negative correlation with the numbers of osteoclasts and expression of marker gene TRAP mRNA and protein kinase K mRNA in osteoclasts. Osteoprotegerin boosted the nitric oxide production and endothelial nitric oxide synthase activity, and osteoprotegerin concentration was positively correlated to the nitric oxide production and endothelial nitric oxide synthase activity. After Raw264.7 cells were in vitro cultured, osteoprotegerin and nitric oxide play a synergic role in inhibiting osteoclasts production and promoting the apoptosis. We speculate that there is osteoprotegerin/endothelial nitric oxide synthase/nitric oxide signal pathway.

Objective To observe the characteristics of vertical cast-off bloodstain pattern by different hitting-tools. Methods The regular hitting tools, a kitchen knife, a dirk, a plane set-hammer and an iron pipe, were selected. At a distance of 30 cmaway fromthe wall, the hitting tool with 5 mL fresh chick-en blood made the cast-off bloodstain fromtop to bottom. Then the holistic distribution characteristics ( length , width and density ) of cast-off bloodstain and morphology characteristics ( length , width and contact angle) of first single cast-off bloodstain were analyzed. Results The distribution length of cast-off bloodstain formed by dirk was minimum( P<0 .05 ) . The distribution width of cast-off bloodstain formed by kitchen knife was minimum(P<0.05). Except the pair of kitchen knife and plane set-hammer, the distribution density between each two tools had statistical differences (P<0.05). The length of first single cast-off bloodstain formed by plane set-hammer was longest compared (P<0.05). The width of first single cast-off bloodstain had statistical differences between kitchen knife and plane set-hammer, and between dirk and plane set-hammer (P<0.05). Conclusion The type of hitting tool could be inferred by the specific characteristics of cast-off bloodstain pattern formed by every specific type of hitting tool in crime scene.

Objective To evaluate the effect of bladder volume change on the doses to normal tissues in cervical cancer patients undergoing external three-dimensional conformal radiotherapy (3DRT)plus 3D conformal brachytherapy (3DCBT).Methods The study included 56 patients with cervical cancer who were admitted to our hospital from 2012 to 2013 and received radical external 3DRT and 3DCBT.During 3DCBT,the doses to 0.1,1.0,and 2.0 cm3(D0.1 cm3,D1.0cm3,and D2.0cm3,respectively) for the rectum,small intestine,sigmoid colon,and bladder under different bladder filling status (empty,50,100,and 150 ml) were compared and analyzed by paired t-test.Results The rectum D0.1cm3 with bladder volumes of 50and 100 ml were significantly reduced compared with that with an empty bladder (P =0.000,0.000).The D0.1 cm3,D1.0cm3,and D2.0cm3 for the small intestine with bladder volumes of 50,100,and 150 ml were significantly reduced compared with those with an empty bladder (P =0.008,0.000,0.000 and 0.000,0.000,0.000 or 0.000,0.000,0.000).The D0.1 cm3,D1.0cm3,and D2.0cm3 for the bladder with bladder volumes of 100 and 150 ml were significantly increased compared with those with an empty bladder (P =0.000,0.000 and 0.000,0.000 or 0.000,0.000).Conclusions The doses to the bladder and small intestine are influenced by different bladder filling status,but the doses to the rectum and sigmoid colon show no significant variation.The increase in bladder volume is helpful in reducing the dose to the small intestine.Without any change in the bladder dose,the bladder volume of 50 ml is more beneficial to reduce the dose to the small intestine than those of 100 and 150 ml.

Objective:To prepare the production of TIGIT-Fc fusion protein using H22 cells stably integrated the gene by lentivirus vector , and to explore the immunoregulatory effect on macrophages by TIGIT-Fc.Methods: TIGIT-Fc fusion gene were constructed by molecular cloning.The fusion gene was then subcloned to plasmids contained the secretion signaling peptide .The secrected TIGIT-Fc fusion gene was inserted into the lentivirus backbone vector.The purified lentivirus vector was the used to infect the murine H22 cell line.TIGIT-Fc protein was purified by protein A column from the ascites of H 22-injected C57BL/6 mice.Macrophages stimulated by lipopolysaccharide ( LPS ) was challenged to TIGIT-Fc treatment or control.Cytokine levels was then detected by ELISA.Results: TIGIT-Fc protein was purified from the ascites of H 22-injected mice.PVR was upregulated in LPS-treated macrophages.IL-10 level was upregulated in TIGIT-Fc treated macrophages.Conclusion: TIGIT-Fc promotes the mature macrophages to secrete anti-inflammatory cytokine IL-10.

Objective Electronic portal imaging devices (EPIDs) are widely used as a replacement of portal films for patient position verification, but the image quality is not always optimal. Because of very low density resolution, the portal imaging is difficult to be used clinically. In this study, several transforming models and the optimization exposure or acquisition conditions were studied for optimization portal imaging, which based on DicomRT platform built by ourselves. Methods 6 MV X-ray from Varian 21EX linac was used to generate portal images by Portal Vision aSiS00 amorphous silicon detector image acquisition system. The density resolution study was based on the number of the lines which could be seen in the image of a special Las Vegas image quality test board. The optimization calculating models were focused on equalization after stretch transforming discrete wavelet transform (DWT) and Butterworth high pass filters. The calculation was performed in Matlab language. Results The optimal numbers of MU, average frames and reset number were 4 - 5,3 - 4 and 2 - 3, respectively. The density resolution of optimized imaging via equalization after stretch transforming, DWT and Butterworth high pass filter transforming was markedly improved. The bone structure could be definitely distinguished. The number of lines distinguished in Las Vegas image via equalization after stretch transforming, DWT and Betterworth high pass filter transforming was 3, 4 and 5, respectivly. Conclusions The proposed transforming systems, including DWT edge detection and Butterworth high pass filter transform, are suitable for improving density resolving power of MV X-ray portal image.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Deoxyadenosines ; pharmacology ; HeLa Cells ; Humans

BACKGROUND:The mechanism and effect of glycogen synthase kinase 3β(GSK-3β) in the differentiation of cardiac stem cel s into cardiomyocytes are stil unclear, although GSK-3βis closely related to the life activities of cel s.
OBJECTIVE:To investigate the changes of GSK-3βexpression in the treatment of myocardial infarction in rats undergoing cardiac stem cel transplantation.
METHODS:The isolation and culture of cardiac stem cel s were performed in 10 neonatal rats. Lentivirus overexpressing GSK-3βor LacZ (control) was constructed and transferred into cardiac stem cel s. Animal model of myocardial infarction was made in 30 Sprague-Dawley rats. Six weeks after model preparation, rat models were assigned into GSK-3β, LacZ or PBS group. GSK-3βor LacZ overexpressing cardiac stem cel solution or PBS in equal volume was injected into the rat myocardium, respectively. Four weeks after transplantation, the cardiac function and myocardial col agen production in rats were detected and compared.
RESULTS AND CONCLUSION:Compared with the other two groups, the left ventricular ejection fraction was significantly higher, and the left ventricular end diastolic diameter was significantly lower in the GSK-3βgroup (P<0.05). Hydroxyproline content, type I col agen mRNA, and type III col agen mRNA expression were significantly lower in the GSK-3βgroup than the other two groups (P<0.05). Findings from Masson staining showed that the content of blue-stained col agen was significantly lower in the GSK-3βgroup than the LacZ group. Moreover, lowest myocardial infarction size was found in the GSK-3βgroup (P<0.05). Al these experimental findings show that GSK-3 overexpression plays a positive role in promoting the therapeutic effect of cardiac stem cel transplantation.

Objective To explore the clinical features of lissencephaly and the detection ofLISI gene.MethodsThe characteristic of clinical features, laboratory examination and gene detection in one case of lissencephaly was retrospectively analyzed. Meanwhile, the related literatures were reviewed.ResultsA 5-month-old female child diagnosed with epilepsy 20 days ago was hospitalized for convulsive seizure more than 30 times in 3 days. The manifestations were eyes staring, and turning upward, cyanosis of lips and face, froth at the mouth, extremities rigidity and loss of consciousness, and the symptoms can spontaneously remitted in 2-3 minutes. Laboratory examination showed that peripheral blood white cell count was 13.67×109/L, hemoglobin 108 g/L, red blood cell count 3.90×1012/L, lymphocyte 10.26×109/L; maocardial enzyme and hepatic and renal function were normal; blood ammonia was 23 μmol/L and lactic acid 2.11 mmol/L. Long-range video EEG showed highly arrhythmia, and frequent partial epilepsy, and sometimes secondary generalized epilepsy. Head MRI showed lissencephaly. The child was treated with oral administration of Keppra 27 mg/(kg·day), Topiramate 6.5 mg/(kg·day), currently no seizure. The detection ofLIS1 gene found that heterozygous mutation of c.232delG, which lead to protein shift mutation (p.E78NfsX25). No mutation was found in her parents.ConclusionsChild with lissencephaly may combine with epilepsy which may cause by mutation inLIS1 gene. And there was no information about point mutation of c.232delG inLIS1 gene being reported at home and abroad so far.

Objective To investigate the application value of BIS monitoring and Ramsay score in the prevention of unplanned tracheal extubation in ICU patients.Methods 93 patients were enrolled in this study,they were divided into the experimental group(47 cases)and the control group(46 cases) using random number method.They received sedation regimens with BIS monitoring and Ramsay score or Ramsay score respectively.Occurrence rate of unplanned extubation was compared between the two groups.Results The occurrence rate of unplanned extubation was significantly lower in the experimental group than that of the control group.Conclusions BIS monitoring and Ramsay score is a suitable ways for the management of sedation of intubated patients.

OBJECTIVE: To observe the characteristics of vertical cast-off bloodstain pattern by different hitting-tools. METHODS: The regular hitting tools, a kitchen knife, a dirk, a plane set-hammer and an iron pipe, were selected. At a distance of 30 cm away from the wall, the hitting tool with 5 mL fresh chicken blood made the cast-off bloodstain from top to bottom. Then the holistic distribution characteristics (length, width and density) of cast-off bloodstain and morphology characteristics (length, width and contact angle) of first single cast-off bloodstain were analyzed. RESULTS: The distribution length of cast-off bloodstain formed by dirk was minimum (P < 0.05). The distribution width of cast-off bloodstain formed by kitchen knife was minimum (P < 0.05). Except the pair of kitchen knife and plane set-hammer, the distribution density between each two tools had statistical differences (P < 0.05). The length of first single cast-off bloodstain formed by plane set-hammer was longest compared (P < 0.05). The width of first single cast-off bloodstain had statistical differences between kitchen knife and plane set-hammer, and between dirk and plane set-hammer (P < 0.05). CONCLUSION The type of hitting tool could be inferred by the specific characteristics of cast-off bloodstain pattern formed by every specific type of hitting tool in crime scene.
Blood Stains ; Computer Simulation ; Crime ; Forensic Ballistics/methods* ; Forensic Medicine/methods* ; Humans