[Evaluation of effect based on different typing methods in Escherichia coli].

Objective: To evaluate the typing and clinical application effect based on clustered regularly interspaced short palindromic repeats (CRISPRs), serotype, and Multilocus Sequence Typing (MLST). Methods: The spacers, serotype and sequence type (ST) were obtained with CRISPRsFinder, SeroTypeFinder and MLST. PCR was used to amplify the CRISPRs, and the spacers were used to predict serotype and ST, then comparing with the serotype and ST. Results: We defined the I-E CRISPR/Cas as CT-Ⅰ, I-F CRISPR/Cas as CT-Ⅱ, and only CRISPR3-4 as CT-Ⅲ. We designated each unique arrangement spacer profile as a unique CRISPRs type. A total of 79 CT types, 76 serotypes, and 66 STs were identified. The CRISPRs typing was the most discriminating, with the Simpson index of 0.936, having the highest correlation with serology with the adjusted Rand index of 0.908. The CRISPRs type could divide the same serotype (ST) into two subtypes [O157∶H7(ST11), O104∶H4(ST678), and O26∶H11(ST21)]. The detection rates of CRISPR1, CRISPR2, CRISPR3, CRISPR4, and CRISPR3-4 were 81.1%, 94.5%, 1.4%, 1.4%, and 4.6%, with the accuracy rate of 95.0% and 100.0% according to the spacers to forecast O157∶H7 (ST11) and ST131. Conclusion: Based on the CRISPRs spacer, this method can be used as an essential molecular typing for E.coli, as it presents a good typing and clinical application effect.

[Clinical application of thoracodorsal artery perforator flaps in the repair of head and neck defects].

Objective: To analyse the clinical application of thoracodorsal artery perforator flaps (TDAPF) in the repair of head and neck defects. Methods: A retrospective review was conducted on 38 patients with oral and maxillofacial head and neck malignant tumors who underwent radical resection of oral and oropharyngeal carcinoma and TDAPF repair in the Department of Oral and Maxillofacial Head and Neck Oncology of the Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from June 2017 to November 2018. Among them, 32 were males and 6 were females, aged 30-74 years. Flap size, vessel pedicle length, diameter and number of perforators, and flap fat thickness were recorded and counted. Elasti Meter and Skin Fibro Meter were applied to measure the skin elasticity and hardness in the donor areas of 4 kinds of skin flaps before the flap preparation. SPSS 19.0 statistical software was used for statistical analysis of the data. Results: All the flaps survived (100%). The mean elasticity of TDAPF [(41.2±12.9) N/m] was significantly lower than that of anterolateral thigh [(77.6±23.3) N/m, χ²=88.89, P<0.05], anterolateral thigh [(62.6±17.7) N/m, χ²=59.99, P<0.05] and or forearm flap [(51.7±8.6) N/m, χ²=37.82, P<0.05]. The hardness of TDAPF [(0.037±0.016) N] was also significantly lower than that of anterolateral femoral [(0.088±0.019) N, F=93.27, P<0.05], anteromedial femoral [(0.059±0.020) N, F=25.71, P<0.05] or forearm flap [(0.062±0.016) N, F=29.11, P<0.05]. Follow-up period ranged from 2 to 14 months. The 38 patients treated with TDAPF had a good recovery of the functions in the recipient areas, and the scars of the donor areas were not obvious after surgery, without serious complications. Conclusion: TDAPF is suitable for reconstruction of head and neck defect, with ductile texture and good recovery of the morphology and function of head and neck.

[Cardioversion efficacy of nifekalan in patients with sustained atrial fibrillation after radiofrequency ablation].

Objective: To evaluate the efficacy and safety of nifekalan (NIF) on cardioversion in atrial fibrillation (AF) patients post radiofrequency ablation, and investigate the relevant factors related to the cardioversion efficacy of NIF. Methods: We screened patients with sustained AF rhythm after radiofrequency ablation between November 2016 and July 2018. Participants were treated with intravenous NIF 0.4 mg/kg within 5-10 minutes after ablation. We observed the adverse reaction, and monitored the rhythm, heart rate, QT interval and QTc interval before the medication and at 5, 10, 20, 120 min after the medication. According to the drug outcome of NIF, patients were divided into conversion group and non-conversion group, related factors affecting conversion efficacy were evaluated using logistic regression analysis. Results: (1)A total of 116 patients were enrolled in the study (63 males and 53 females, mean age was (64±18) years). Among them, 72 patients were converted to sinus rhythm, and the overall successful rate was 62.1%. There were 84 patients with persistent AF, of which 50 cases (59.2%) were restored to sinus rhythm. There were 32 patients with paroxysmal AF, 22 cases (68.8%) of them were restored to sinus rhythm. The conversion time was 1.5 to 12 (6.8±3.4)min. (2) In 116 patients, the QT interval and QTc interval were significantly longer after medication than before the drug administration (P<0.01), and peaked at about 10th min, and restored to the level before drug administration at about 120th min. (3) There were 8 cases of bradycardia (6.9%), 3 cases of frequent and short ventricular tachycardia (2.6%). (4) The duration of atrial fibrillation was shorter and left atrial diameter was smaller in the cardioversion group than in the non-cardioversion group (both P<0.05). There were no significant differences in gender, disease history, atrial fibrillation type and structural heart disease between the two groups (P>0.05). (5) Multifactorial logistic regression analysis showed that the duration of atrial fibrillation (OR=0.980, 95%CI 0.966-0.994, P=0.004) and the left atrial diameter (OR=0.888, 95%CI 0.814-0.967, P=0.007) were the factors that influence the cardioversion efficacy of NIF on atrial fibrillation post ablation. Conclusions: The total effective rate of NIF was 62.1% in patients witrh sustained AF post radiofrequency ablation, was 68.8% in patients with paroxysmal AF. Besides, NIF has the advantage of short conversion time and few adverse reactions. Left atrium diameter and AF duration were relevant factors that influence the efficacy of NIF of cardioversion in patients with sustained AF after radiofrequency ablation.

Role of a two-step suture in the prevention of postoperative transoral salivary fistulas during reconstruction of the oral cavity.

Transoral salivary fistulas are one of the most serious postoperative complications after operations for oral cancer, and we propose a new, two-step suture method to avoid them. From January 2005 to September 2017, 240 patients were recruited at the Shanghai Ninth People's Hospital and divided into experimental (n=89) or control (n=151) groups. The experimental group was treated by a two-step suture technique, while the control group had conventional sutures. Statistical differences were assessed using the chi squared and t tests, as appropriate. Only two patients developed transoral salivary fistulas in the experimental group, while in the control group there were 14 (9%). The incidence of fistulas in the experimental group was significantly lower than that in the control group (p=0.035). Regression analysis showed that there was a significant correlation between the groups and the incidence of salivary fistulas (p=0.032). The two-step suture technique is safe, effective, and easy to learn, and could reduce the incidence of postoperative salivary fistulas.

MiR-181d promotes steroid-induced osteonecrosis of the femoral head by targeting SMAD3 to inhibit osteogenic differentiation of hBMSCs.

OBJECTIVE: We aimed to investigate whether miR-181d may be involved in steroid-induced osteonecrosis of the femoral head (ONFH) and its underlying mechanism. PATIENTS AND METHODS: Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-181d in bone marrow of 5 cases of steroid-induced femoral head necrosis and 5 cases of femoral head necrosis secondary to femoral neck fractures. Marrow-derived mesenchymal stem cells (MSCs) were obtained from bone marrow samples and identified. Subsequently, the effects of miR-181d on osteogenic differentiation were evaluated by alizarin red staining and alkaline phosphatase. Meanwhile, qRT-PCR was performed to detect the levels of osteoblast-specific genes. RESULTS: The expression of miR-181d in the bone marrow of patients with steroid-induced osteonecrosis of the femoral head was significantly higher than that in the control group. When the expression of miR-181d in MSCs was upregulated, the ALP staining became lighter and the number of calcified nodules, as well as the expression of osteoblast-specific genes, decreased significantly. Meanwhile, the opposite results were observed when miR-181d expression was inhibited. Western blot and luciferase reporting assay proved that miR-181d could negatively regulate the expression of SMAD3. CONCLUSIONS: MiR-181d can inhibit the differentiation of hBMSCs into osteoblasts by regulating the expression of SMAD3.

[Recurrent syncope related to catecholaminergic polymorphic ventricular tachycardia due to de novo RyR2-R2401H mutation].

Objective: To explore the clinical and molecular genetic features of a Chinese patient with catecholaminergic polymorphic ventricular tachycardia (CPVT). Methods: Clinical data including resting electrocardiography, echocardiography and treadmill exercise testing of a patient with CPVT admitted to our department in March 2013 were analyzed, and the peripheral venous blood samples of the patient and his family members and 400 ethnicity-matched healthy controls were obtained. All exons and exon-intron boundaries of the six CPVT-related genes including RYR2, CASQ2, TRDN, CALM1, KCNJ2 and ANKB were sequenced to detect the variants related to CPVT. The relationship between the genotypes and phenotypes was analyzed to direct the target therapy. Results: Recurrent syncope induced either by exercise or extreme frightened fear was observed in this patient. There was no positive family history of syncope or sudden death. The resting electrocardiography and echocardiography of the patient were normal, while the exercise testing revealed bidirectional and polymorphic ventricular tachycardia. A cardiac ryanodine receptor gene mutation (R2401H) was identified in this patient, while this mutation was absent in his parents and sister and 400 controls. No variant was detected in the remaining five candidate genes. Treatment with high dose of metoprolol succinate (118.75 mg/d) was effective and patient was free of syncopal attack during the 2 years follow-up. Conclusion: This is the first report on RyR2-R2401H mutation in Chinese patient with CPVT, and high dose of metoptolol is the effective therapy option for CPVT related to RyR2 mutation.

1H NMR Metabolomics Study of Spleen from C57BL/6 Mice Exposed to Gamma Radiation.

Due to the potential risk of accidental exposure to gamma radiation, it's critical to identify the biomarkers of radiation exposed creatures. In the present study, NMR based metabolomics combined with multivariate data analysis to evaluate the metabolites changed in the C57BL/6 mouse spleen after 4 days whole body exposure to 3.0 Gy and 7.8 Gy gamma radiations. Principal component analysis (PCA) and orthogonal projection to latent structures analysis (OPLS) are employed for classification and identification potential biomarkers associated with gamma irradiation. Two different strategies for NMR spectral data reduction (i.e., spectral binning and spectral deconvolution) are combined with normalize to constant sum and unit weight before multivariate data analysis, respectively. The combination of spectral deconvolution and normalization to unit weight is the best way for identifying discriminatory metabolites between the irradiation and control groups. Normalized to the constant sum may achieve some pseudo biomarkers. PCA and OPLS results shown that the exposed groups can be well separated from the control group. Leucine, 2-aminobutyrate, valine, lactate, arginine, glutathione, 2-oxoglutarate, creatine, tyrosine, phenylalanine, π-methylhistidine, taurine, myoinositol, glycerol and uracil are significantly elevated while ADP is decreased significantly. These significantly changed metabolites are associated with multiple metabolic pathways and may be potential biomarkers in the spleen exposed to gamma irradiation.

The effect of estrogen-related receptor α on the regulation of angiogenesis after spinal cord injury.

Estrogen receptor-related receptor-α (ERRα) is an orphan member of the nuclear receptor superfamily that interacts with peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) to stimulate vascular endothelial growth factor (VEGF) expression and angiogenesis in a hypoxia-inducible factor-1α-independent pathway. Although it is not regulated by any natural ligand, the action of ERRα can be blocked by the synthetic molecule XCT790. In the present study, Sprague-Dawley rats were randomly allocated to a sham group, injury-saline group or injury-XCT90 group. A modified Allen's weight-drop method was applied to induce the acute traumatic spinal cord injury (SCI) model in these rats, and an injection of XCT790 was administered every 24h, starting half an hour after the SCI contusion. Histological analyses revealed that XCT790 significantly aggravated tissue damage and decreased the number of ERRα-positive cells at 1, 3 and 7 days after SCI. Western blot and quantitative real-time polymerase chain reaction (qRT-PCR) analyses also indicated that XCT790 dramatically repressed the expression of ERRα, thus reducing the expression of VEGF and angiopoietin-2 (Ang-2) throughout the duration of the experiment, but the expression of PGC-1α was not affected. Immunofluorescence analyses indicated that vascular density and endothelial cell proliferation were decreased in the injury-XCT90 group compared with the injury-saline group. These results suggest that ERRα is involved in mediating angiogenesis after SCI in the rat traumatic SCI model.

Does cross-linked polyethylene decrease the revision rate of total hip arthroplasty compared with conventional polyethylene? A meta-analysis.

BACKGROUND: Although cross-linked polyethylene is resistant to wear in comparison to conventional polyethylene, it remains unknown whether it can decrease the wear-related revision rate of total hip arthroplasty. OBJECTIVES: To determine whether cross-linked polyethylene decreases the wear-related revision rate of total hip arthroplasty compared with conventional polyethylene. DATA SOURCES: Electronic databases, including PubMed, EMBASE, and the Cochrane Central Register of Controlled Trials, were queried from inception to July 6, 2013. STUDY SELECTION: Randomized controlled trials (RCTs) comparing cross-linked polyethylene with conventional polyethylene were included. In addition, the standard 28-mm femoral head was used, and follow-up was performed for a minimum of 5 years. The primary outcome assessed was wear-related revision. The secondary outcome measures evaluated were the incidence of osteolysis, the linear wear rate, and the linear head penetration. DATA SYNTHESIS: The Cochrane Collaboration's tool for assessing the risk of bias was used for quality assessment. Data from eligible studies were pooled using a random effects model. RESULTS: Eight studies involving 735 patients were included in this study. Meta-analysis showed there was no significant difference between cross-linked and conventional polyethylene group in terms of osteolysis or wear-related revision. The pooled mean differences were significantly less for the linear wear rate and linear head penetration for cross-linked polyethylene than for conventional polyethylene. LIMITATIONS: The studies differed with respect to the cross-linked liner brands, manufacturing processes, and radiological evaluation methods. Moreover, the follow-up periods of the RCTs were not long enough. CONCLUSIONS: The current limited evidence suggests that cross-linked polyethylene significantly reduced the radiological wear compared with conventional polyethylene at midterm follow-up periods. However, there is no evidence that cross-linked polyethylene had an advantage over conventional polyethylene in terms of reducing osteolysis or wear-related revision. Nevertheless, future long-term RCTs on this topic are needed. KEY FINDINGS: Cross-linked polyethylene significantly reduced radiological wear but not osteolysis or wear-related revision in comparison to conventional polyethylene at midterm follow-up periods. LEVEL OF EVIDENCE: Level I, systematic review of level I studies.

Metastatic Melanoma Induced Metabolic Changes in C57BL/6J Mouse Stomach Measured by 1H NMR Spectroscopy.

Melanoma is a malignant tumor of melanocytes with high capability of invasion and rapid metastasis to other organs. Malignant melanoma is the most common metastatic malignancy found in Gastrointestinal Tract (GI). In this work, the 1H NMR-based metabolomics approach is used to investigate the metabolite profile differences of stomach tissue extracts of metastatic B16-F10 melanoma and control groups in C57BL/6J mouse and to search for specific metabolite biomarker candidates. Principal Component Analysis (PCA), an unsupervised multivariate data analysis method, is used to detect possible outliers, while Orthogonal Projection to Latent Structure (OPLS), a supervised multivariate data analysis method, is employed to evaluate important metabolites responsible for discriminating the control and the melanoma groups. Both PCA and OPLS results reveal that the melanoma group can be well separated from its control group. Among the 50 identified metabolites, it is found that the concentrations of 19 metabolites are significantly changed with the levels of O-phosphocholine and hypoxanthine down-regulated while the levels of isoleucine, leucine, valine, isobutyrate, threonine, cadaverine, alanine, glutamate, glutamine, methionine, citrate, asparagine, tryptophan, glycine, serine, uracil, and formate up-regulated in the melanoma group. These significantly changed metabolites are associated with multiple biological pathways and may be potential biomarkers for metastatic melanoma in stomach.

In vivo measurement of the subchondral bone thickness of lumbar facet joint using magnetic resonance imaging.

OBJECTIVE: To measure in vivo thicknesses of the facet joint subchondral bone across genders, age groups, with or without low back pain symptom groups and spinal levels. METHODS: Lumbar (L1-L2 to L5-S1) magnetic resonance (MR) imaging was performed in 81 subjects (41 males and 40 females, mean age 37.6 years). Thicknesses of the subchondral bone were measured in 1620 facet joints using the MR images with custom-written image processing algorithms together with a multi-threshold segmentation technique using each facet joint's middle axial-slice. This method was validated with 12 cadaver facet joints, scanned with both MR and micro-computed tomography images. RESULTS: An overall average thickness value for the 1620 analyzed joints was measured as 1.56±0.01 mm. The subchondral bone thickness values showed significant increases with successive lower spinal levels in the subjects without low back pain. The facet joint subchondral bone thickness in asymptomatic females was much smaller than in asymptomatic males. Mean subchondral bone thickness in the superior facet was greater than that in the inferior facet in both female and male asymptomatic subjects. CONCLUSIONS: This study is the first to quantitatively show subchondral bone thickness using a validated MR-based technique. The subchondral bone thickness was greater in asymptomatic males and increased with each successive lower spinal level. These findings may suggest that the subchondral bone thickness increases with loading. Furthermore, the superior facet subchondral bone was thicker than the inferior facet in all cases regardless of gender, age or spinal level in the subjects without low back pain. More research is needed to link subchondral bone microstructure to facet joint kinematics and spinal loads.

Stimulation of the sacral nerve reduces gut bacterial translocation and endotoxemia caused by acute spinal cord injury in rabbits.

Study design:Disturbance of gastrointestinal motility following acute spinal cord injury complicated with paraplegia can lead to bacterial overgrowth in the gastrointestinal tract, and increase the incidence of bacterial translocation. Experiments in a New Zealand rabbit model of acute spinal cord injury were performed.Objective:This study was designed to determine if the electrical stimulation of the sacral nerve prevents gut-origin bacterial translocation and endotoxemia in an animal model of acute spinal cord injury.Settings:Xiangya Hospital, Central South University, Hunan, PR China.Methods:New Zealand rabbits were divided into three groups: Control group, Paraplegic Vehicle group without sacral nerve stimulation and Paraplegic Experiment group with sacral nerve stimulation. Blood and multiple organs were collected for bacterial cultures and endotoxin determination at 24, 48 and 72 h following spinal cord injury. The histology and ultra-structural features of the organs were studied.Results:Bacterial translocation and endotoxemia were observed in all animals with acute spinal cord injury. Sacral nerve stimulation increased defecation, decreased endotoxin levels and bacterial translocation and improved the morphology of the organs.Conclusion:After acute spinal cord injury, stimulation of the sacral nerve reduced gut bacterial translocation and endotoxemia.Spinal Cord advance online publication, 6 April 2010; doi:10.1038/sc.2010.35.

Accumulation and acute toxicity of silver in Potamogeton crispus L.

In the present study, Potamogeton crispus L. plants exposed to various concentrations of silver (Ag) (5, 10, 15, and 20 microM) for 5d were investigated to determine the accumulating potential of Ag and its influence on nutrient elements, chlorophyll pigments and fluorescence, various antioxidant enzymes and compounds, adenosine triphosphate (ATP), protein content and ultrastructure. The accumulation of Ag was found to increase in a concentration dependent manner with a maximum of 29.3 microg g(-1) at 20 microM. The nutrient elements (except Ca), photosynthetic pigments, chlorophyll a fluorescence parameters (Fo, Fv, Fv/Fm, Fv/Fo), malondialdehyde (MDA), ATP, peroxidase (POD) activity, ascorbate (AsA), reduced glutathione (GSH) and protein contents decreased significantly as concentration of Ag augmented. In contrast, an induction in SOD activity was recorded, while an initial rise in Ca content and CAT activity was followed by subsequent decline. Morphological symptoms of senescence phenomena such as chlorosis and damage of chloroplasts and mitochondria were observed even at the lowest concentration of Ag, which suggested that Ag hastened the senescence of the tested plants. The loss of nutrients and chlorophyll content and damage of chloroplasts were associated with disturbances in photosynthetic capacity as indicated by the quenching of chlorophyll a fluorescence. Decreased chlorophyll and protein contents suggest oxidative stress induced by Ag. In addition, both the reduction of ATP and the damage to the ultrastructure of organelles were indicative of general disarray in the cellular functions exerted by Ag.

Diamond anvil cell radial x-ray diffraction program at the National Synchrotron Light Source.

During the past decade, the radial x-ray diffraction method using a diamond anvil cell (DAC) has been developed at the X17C beamline of the National Synchrotron Light Source. The detailed experimental procedure used with energy dispersive x-ray diffraction is described. The advantages and limitations of using the energy dispersive method for DAC radial diffraction studies are also discussed. The results for FeO at 135 GPa and other radial diffraction experiments performed at X17C are discussed in this report.

A novel dipolar dephasing method for the slow magic angle turning experiment.

Complete suppression of the resonances from protonated carbons in a slow magic angle spinning experiment can be achieved using five dipolar dephasing (Five-DD) periods distributed in one rotor period. This produces a spectrum containing only the spinning sidebands (SSB) from the nonprotonated carbons. It is shown that the SSB patterns corresponding to the nonprotonated carbons are not distorted over a wide range of dipolar dephasing times. Hence, this method can be used to obtain reliable principal values of the chemical shift tensors for each nonprotonated carbon. The Five-DD method can be readily incorporated into isotropic-anisotropic 2D experiments such as FIREMAT and 2D-PASS to facilitate the measurement of the (13)C chemical shift tensors in complex systems.

High-resolution (1)H NMR spectroscopy in organs and tissues using slow magic angle spinning.

It is shown that high-resolution (1)H NMR spectra of intact excised tissues and organs can be obtained by rotating the sample slowly about an axis at the magic angle of 54 degrees 44' with the external magnetic field. In this way tissue and cellular damage invoked by standard magic angle spinning (MAS) experiments, where spinning speeds of several kHz are typically employed, are minimized. Special RF pulse sequences, developed originally in solid state NMR, can be used to produce a spinning sideband-free isotropic spectrum. In this article the first results are shown of the brain, heart, liver, gluteus muscle, and kidney excised from mice using the 2D-phase-altered spinning sidebands (PASS) technique and employing MAS spinning speeds of 43-125 Hz. It was found that with slow sample spinning similar, and in some cases even better, spectral resolutions are obtained as compared with fast MAS.

[Observation on effect of radiotherapy and antike capsule combination therapy in treating nasopharyngeal cancer patients].

OBJECTIVE: To compare the effect of radiotherapy (RT) combined with Antike capsule (AC) and RT alone in treating nasopharyngeal cancer (NPC) patients. METHODS: Eighty-nine patients with pathologically confirmed NPC (stage II-IV) were randomly divided into two groups: group A (46 cases) were treated with RT, receiving 65-70 Gy/6.5-7 weeks to nasopharynx region and the same dosage to neck region, and AC was given in combination. Group B (43 cases) received the same RT alone. The total dosis of RT for complete remission (CR) of primary nasopharyngeal tumor and neck lymph nodes, the CR rate and the changes of peripheral NK cell, T lymphocyte subsets in the two groups were compared. RESULTS: The total dosis of RT for CR in group A and B were 41.6 +/- 8.9 Gy vs 50.7 +/- 9.2 Gy for primary nasopharyngeal tumor, P < 0.05 and 47.4 +/- 10.3 Gy vs 56.2 +/- 9.7 Gy for neck lymph nodes, P < 0.05. The CR rate of primary nasopharyngeal tumor in group A and B were 93.5% and 88.4% respectively, P < 0.05. The activity of NK cell as well as T3, T4 in peripheral blood increased significantly in the group A after treatment, P < 0.05, while in group B, T3, T4 lowered significantly, P < 0.05. CONCLUSION: RT combined with AC could be helpful in elevating and promoting the remission rate of primary tumor and neck lymph nodes, and AC has some effects in improving the immune function and general condition in NPC patients during RT.

[Expressions of p-MAPK, cyclin D1, p53 protein and their relationship in osteosarcoma].

OBJECTIVE: To detect the expressions of p-MAPK, cyclin D1 and p53 protein, and investigate their relationship in osteosarcomas. METHODS: SP immunohistochemical technique was used to detect the expressions of of p-MAPK, cyclin D1 and p53 protein among the 58 osteosarcomas and 14 osteoid osteomas samples. RESULTS: The positive rates of p-MAPK, cyclin D1 and p53 protein were 77.6% (45/58), 60.3% (35/58) and 44.8% (26/58) respectively in 58 osteosarcomas, and 21.4% (3/14), 21.4% (3/14) and 7.1% (1/14) in 14 osteoid osteomas. Their positive rates and expressive intensities in osteosarcomas were higher than those in osteiod osteomas. Their positive correlation was significantly observed between p-MAPK and cyclin D1 proteins (P < 0.01), and wasn't observed between p-MAPK and p53 proteins (P < 0.05) in osteosarcomas. CONCLUSIONS: MAPK phosporylation may be one of causes for activation of cyclin D1, which play an important role in osteosarcogenesis. The mutation of p53 oncogene may not result in osteosarcogenesis by MAPK signaling pathway. Detecting p-MAPK by immunohistochemistry may be one of diagnostic indexes for osteosarcoma and benign osteoma.

A high-resolution (13)C 3D CSA-CSA-CSA correlation experiment by means of magic angle turning.

It is shown in this paper that a previously reported 90 degrees sample flipping (13)C 2D CSA-CSA correlation experiment may be carried out alternatively by employing constant slow sample rotation about the magic angle axis and by synchronizing the read pulse to 13 of the rotor cycle. A high-resolution 3D CSA-CSA-CSA correlation experiment based on the magic angle turning technique is reported in which the conventional 90 degrees 2D CSA-CSA powder pattern for each carbon in a system containing a number of inequivalent carbons may be separated according to the isotropic chemical shift value. The technique is demonstrated on 1,2,3-trimethoxybenzene in which all of the overlapping powder patterns that cannot be segregated by the 2D CSA-CSA experiment are resolved successfully by the 3D CSA-CSA-CSA experiment, including even the two methoxy groups (M(1) and M(3)) whose isotropic shifts, confirmed by high-speed MAS, are separated by only 1 ppm. A difference of 4 ppm in the principal value component (delta(33)) between M(1) and M(3) is readily obtained.