Objective To compare two separation medium of isolation of adult rat cardiomyocytes , and to observe the characteristics of excitation-contraction coupling of cardiomyocytes .Methods The isolated adult rat heart was hanged on to the Langendorff apparatus for aortic counter-current perfusion and collagenase digestion using two different separation medium.The single cardiomyocytes were cultured and infected with adenovirus . The morphological features of cardiomyocytes were observed with microscope and fluorescent microscope . The shortening-re-lengthening features of sarcomere and the intake-discharge features of calcium were simultaneously recorded by IonOptix equipment .Results 70%rod-shaped with clear-striation adult rat cardiomyocytes could be obtained with the stated two separation medium and cultured in serum-free medium for more than 7 days.GFP could express more than 7 days when the cardiomyocytes were infected with adenovirus .Cardiomyocytes obtained by the first separation medium could not contract with the electrical stimulation, while cardiomyoctyes obtained by the second separation medium could be used for the detection of excitation -contraction coupling .The shortening fraction of sarcomere was 11.61%±2.15% and the relaxing time was ( 0.177 ± 0.031) s.The amplitude of calcium transient was 30.79% ±9.74 % and the decaying time of calcium transient was (0.300 ±0.074) s.Conclusion With the stated two separation medium , adult rat cardiomyocytes can be well isolated , cultured and infected with adenovirus .The second separation medium can be used for the detection of excitation-contraction coupling characteristics .

Objective To evaluate the clinical results of the Ilizarov technique for the treatment of the complex rigid talipes equinovarus deformities.Methods From July 2005 to July 2011,28 patients (41 feet) with rigid talipes equinovarus deformities which had been corrected with the Ilizarov technique were retrospectively analyzed,including 18 males (26 feet) and 10 females (15 feet) with an average age of 15.3 years.According to the classification system proposed by Diméglio,31 feet were categorized as grade Ⅲ,and 10 as grade Ⅳ.We performed corrections with a soft tissue release in 23 feet,and with a limited osteotomy in 18,and then a Ilizarov external fixator was applied.Anteroposterior and lateral X-rays were taken to compare the pre and postoperative data in terms of the angle of plantarflexion and dorsiflexion,the range of motion of the ankle joint,radiological measurements of the talocalcaneal angle.Results All the 28 patients achieved an outpatient follow-up,with an average of 25 months.All patients achieved a plantigrade foot with an almost normal appearance as the fixator was removed after applied for an average of 5.1 months (range,2-14).At the preoperative and final follow-up respectively,the angle of dorsiflexion of the foot was -45.0°±12.0° and 9.5°±5.5°,the angle of plantarflexion was 67.0°±14.0° and 45.5°±7.8°,talocalcaneal angle was 6.5°±4.5° and 22.5°±5.5° in anteroposterior radiograph and 5.5°±11.0° and 40.6°±8.5° in lateral radiograph.Spastic ischemia occurred in one foot and relieved by a slower distraction rate.Wire-hole infections occurred in 5 feet and treated by dressing changs,wire tract altering and antibiotic therapy,finally the infections were controlled.Deformity relapsed in one foot three months after the device was removed,then corrected with an additional fixator application and has not recurred till the final follow-up.Toe contracture and residual deformity occurred in 5 feet and 3 feet,respectively.Conclusion The Ilizarov technique is an effective method for correction of complex rigid talipes equinovarus deformities,with which the appearance and function of the foot could be kept as much as possible,and without impact on food development.

Objective To investigate the risk factors affecting the malignancy of amorphous calcifications in the breast and to establish a predictive nomogram.Methods Patients with amorphous calcifications detected by mammography were retrospectively collected,clinical data were obtained from electronic medical record(EMR),and the mammographic features of the patients were assessed by diagnostic physicians.The risk factors affecting the malignancy of amorphous calcifications were analyzed to develop a predictive model and to assess the performance of the model.Results A total of 153 amorphous calcifications in 144 patients were included in the study,and the overall malignancy rate of calcifications was 20.92％.Patient's age ≥45 years,linear distribution of calcifications,unilateral single or unilateral multiple calcifications,and a larger maximum ratio of calcification extent all predicted a higher probability of malignancy,establishing a nomogram based on these 4 risk factors,with a 3.65％predicted probability of malig-nancy as the cut-off,33.99％(52/153)of patients were allowed to be spared biopsy.Conclusion Patient's age and the distribution,number,and maximum ratio of calcifications may be the risk predictors of malignancy for amorphous calcifications,with nomogram con-struction for distinguishing benignity from malignancy of amorphous calcifications via combining with mammographic features and clinical data.

Objective To observe the value of breast imaging reporting and data system(BI-RADS)classification combined with mammography(MG)radiomics for differentiating breast benign and malignant amorphous calcification lesions.Methods A total of 217 breast amorphous calcification lesions in 206 female patients,including 43 malignant and 174 benign ones were retrospectively enrolled and randomly divided into training set(n=151)and validation set(n=66)at the ratio of 7∶3.Then radiomics features were screened based on breast MG images.BI-RADS model,radiomics model and combined model were established,and the value of each model for distinguishing benign and malignant lesions was observed.Results Eight optimal radiomics features were selected.The area under the curve(AUC)of BI-RADS model,radiomics model and combined model for differentiating breast benign and malignant amorphous calcification lesions was 0.821,0.763 and 0.897 in training set,0.800,0.746 and 0.893 in validation set,respectively,AUC of combined model was significantly higher than that of the other two(both P＜0.05).Conclusion BI-RADS classification combined with MG radiomics was helpful for differential diagnosis of breast benign and malignant amorphous calcification lesions,with efficacy higher than that of each alone.

Influenza is one of the most common infections threatening public health worldwide and is caused by the influenza virus. Rapid emergence of drug resistance has led to an urgent need to develop new anti-influenza inhibitors. In this study we established a 293T cell line that constitutively synthesizes a virus-based negative strand RNA, which expresses Gaussia luciferase upon influenza A virus infection. Using this cell line, an assay was developed and optimized to search for inhibitors of influenza virus replication. Biochemical studies and statistical analyses presented herein demonstrate the sensitivity and reproducibility of the assay in a high-throughput format (Z' factor value>0.8). A pilot screening provides further evidence for validation of the assay. Taken together, this work provides a simple, convenient, and reliable HTS assay to identify compounds with anti-influenza activity.

ObjectiveTo explore the clinical efficacy of Huanglian Jiedutang as an adjunctive treatment for acute cerebral infarction complicated with gastric motility disorder. MethodSixty patients with acute cerebral infarction complicated with gastric motility disorder with fire toxin syndrome were randomly divided into a western medicine control group （control group） and a traditional Chinese medicine （TCM） combined treatment group （observation group）， with 30 cases in each group. The control group received basic treatment for cerebral infarction and relevant western medical symptomatic treatment based on the patients' gastrointestinal symptoms. The observation group received Huanglian Jiedutang in addition to the treatment provided to the control group. The treatment course was 7 days. Neurological deficit scores and gastrointestinal dysfunction scores were assessed in both groups before treatment and on the 4^th and 7^th days of treatment. Gastrointestinal electrographic parameters， serum citrulline （CIT）， and motilin （MTL） levels were measured in both groups before treatment and on the 7^th day of treatment. Clinical efficacy was compared between the two groups. ResultCompared with the baseline in both groups， the neurological deficit scores and gastrointestinal dysfunction scores were significantly reduced on the 4th and 7^th days of treatment （P<0.05）. The reductions in these scores were more significant on the 7^th day compared with those on the 4^th day of treatment （P<0.05）. On the 4^th and 7^th days of treatment， the observation group showed a significantly greater reduction in neurological deficit scores and gastrointestinal dysfunction scores compared with the control group （P<0.05）. On the 7th day of treatment， compared with the baseline， both groups showed a significant increase in gastric antral and gastric body electric wave amplitudes as well as serum CIT and MTL levels （P<0.05）， and there were no statistically significant differences in the frequency of gastric antral and gastric body electric waves. On the 7th day of treatment， compared with the control group， the observation group had a significant increase in gastric antral and gastric body electric wave amplitudes as well as serum CIT and MTL levels （P<0.05）， and there were no statistically significant differences in the frequency of gastric antral and gastric body electric waves. After 7 days of treatment， the total effective rate in the observation group was 90.00% （27/30）， higher than 76.67% （23/30） in the control group， but the difference was not statistically significant. ConclusionAdjunctive treatment with Huanglian Jiedutang can effectively improve the symptoms of neurological function impairment and gastrointestinal dysfunction in patients with acute cerebral infarction complicated with gastric motility disorder， increase gastric antral and gastric body electric wave amplitudes， improve gastric motility disorder， and increase serum CIT and MTL levels， thereby improving the imbalanced secretion function of the gastrointestinal tract.