Objective To study the effects of mechanical strain on the proliferation of the human pulmonary epithelial cell and the redistribution of its membrane receptors, integrins ? 5 and ? 1. Methods A cyclic strain unit in vitro was designed. The cellular proliferative index was measured by flow cytometry and the redistribution of ? 5 and ? 1 integrins was analyzed in human pulmonary epithelial cell line H727 by laser confocal microscopy. Results The cellular proliferative index reduced significantly after cells were subjected to 15% elongation at frequencies of 20 cycles/min or 40 cycles/min for 24 h. In human pulmonary epithelial H727 cells, ? 5 and ? 1 integrins transferred from the apical layer to the basal layer and formed an adhesion plaque after 24 h exposure to 15% elongation at frequency of 40 cycles/min. Conclusion The results suggest that ? 5 and ? 1 integrins in pulmonary epithelial cells may play an important role in the transduction of mechanical stress.

Objective To observe the survival,conversion and function preservation of mesenchymal stem cells(MSC) after transplantation.Methods MSCs from male Sprague-Dawley(SD) rats were isolated and cultured,then were transplanted to bone defect of female SD rats.Tissues were obtained at defferent periolds.Hybridization in situ was performed on the tissues using Y chromosome specific DNA probe labeled with digoxin to observe the transplanted cells on distribution,quantity and function.Results Allogeneic MSCs could survive and proliferate in the bone defect of recipients and formed bony callus with large quantity and good quality.After 90 days of graft,stem cells still survived and distributed in marrow and newly formed bone tissue.Conclusion After transplantation,allogeneic MSCs have a long-term surviving in marrow and new bone tissue and maintain the characteristic of ossification.

Objective To analyze the protein expression and subcellular distribution of mechanogrowth factor (MGF) in ostcoblasts under stretch stimulation. Methods Cyclic stretching was applied to osteohlasts by a mechanical stretching device. The whole-cell proteins were extracted from controlled and stretched osteoblasts for detecting the protcin expression level of MGF by Western blot and observing the intracellular distribution of MGF by fluorescent immunocytological method. Results Western blot showed significant increase of expression of MGF in osteoblasts under stimulation of cyclic stretching. The level of protein was increased by four folds after 12-hour stretching of osteohlasts, and then declined sharply. Immunofluorescence analysis showed that MGF was mainly distributed in the nuclei of osteoblasts. ConcinsionsUnder the cyclic stimulation, the expression of MGF reaches a short period of peak in osteoblasts, which may be related to the injury of osteoblasts caused by stretching. MGF is mainly distributed in the nuclei of osteoblasts, indicating that MGF may contain nuclear localization signal and modulate the expression of relative genes.

Objective To explore the clinical results of the arthroscopic acromioplasty in the rotator cuff repair.Methods Sixty-five patients (42 males and 23 females) to receive suture bridge repair under the arthroscopy between May 2012 and May 2014 were selected and randomly divided into an experimental group and a control group.The experimental group was given suture bridge repair with acromioplasty,while the control group underwent the suture bridge repair without acromioplasty.The time of operation was recorded.The range of motion (ROM) of the shoulder,the UCLA shoulder scoring system (ULCA) score,American orthopedic surgeon scoring system (ASES) score,the visual analogue scale (VAS) score and Constant score were observed 3 and 12 months after the operation.Results There were no significant differences in age,sex and the affected side between the two groups.Significant differences were found between the two groups in the duration of the operation (t=-18.5,P＜0.05).There were no significant differences in the ROM and the ULCA,ASES,VAS and constant scores of the shoulder 3 and 12 months after the operation.Conclusion No significant differences were found in the ROM and the shoulder function scoring between giving acromioplasty under the arthroscopy or not.However,the operation time of undergoing acromioplasty under the arthroscopy was longer.

Objective:To investigate the effects of icaritin(ICT)on the proliferation and osteogenic differentiation of rat bone mar-row stromal cells(rBMSCs).Methods:rBMSCs were cultured from the bone marrow of SD rats and identified by multilineage differ-entiation assays.3,6 and 9 days after the treatment of rBMSCs of passage 4 by ICT at 1 0 -9 ,1 0 -8 ,1 0 -7 ,1 0 -6 and 1 0 -5 mol/L re-spectively,the proliferation and differentiation of the cells were examined by cck-8 and alkaline phosphatase (ALP)activity assay kit respectively.The calcium nodule formation was observed by alizarin red(AR)staining 21 days after 1 0 -9 mol/L ICT treatment. Results:Primary rBMSCs showed the typical spindle-like shape with attachment growth.rBMSCs could be induced to osteogenic and adipogenic differentiation.The proliferation of rBMSCs was inhibited but ALP activity was enhanced by ICT.1 0 -9 mol/L ICT in-cresed calcium nodule formation.Conclusion:ICT can dose-dependently inhibit the proliferation,but promote the osteogenic differ-entiation of rBMSCs.

Objective:To investigate the correlation between the breast cancer MRI schedule of reinforcement and the shrinkage pattern of tumor after neo-adjuvant chemotherapy, and its clinical significance in the guidance of formulating operation plan. Methods:Dynamic contrast-enhanced MRI scan was performed before chemotherapy and before surgery after a whole-range N-Acety-L-Cysteine (NAC) treatment in 55 patients with loco-regionally advanced breast cancer who received the neo-adjuvant chemotherapy. MRI sched-ule of reinforcement and the shrinkage pattern of tumor after neo-adjuvant chemotherapy were obtained in the treatment, and the corre-lation between the two was analyzed. Results:Of the 55 patients, the unilateral breast mass was found in 54 and the bilateral lesion in 1. There were 56 neo-plastic foci in these patients. The mass-like enhanced image was seen in 24 of the total cases (43%), of which 23 presented with a centripetal shrinkage, 1 with an annular decline, (P<0.01). Multifocal mass-like enhancement image was found in 13 of the total cases, (23%), of which 2 centripetal shrinkages were the singular mass, 11 remained a multifocal lesion after the tumor shrinkage (P<0.01). The mass with peripheral non-tumor-like enhancement image was seen in 8 of the total cases (14%), of which 4 showed a centripetal shrinkage and another 4 a honeycombed multifocal decline (P>0.05). There were 11 of the total cases with non-tu-mor-like enhancement (20%), in which 4 assumed a centripetal shrinkage and 7 a honeycombed multifocal shrinkage (P<0.01). Conclu-sion:The tumor shrinking pattern and its accurate radiological image evaluation are the keys to the selection of breast-conserving sur-gery and the control of local recurrence after treatment of NAC regimen. We can predict the shrinking pattern through the type of the le-sion on baseline before NAC, which is important for the patients and surgeon to get a reasonable expectation in the subsequent treat-ments.

Objective:To investigate the value of artificial intelligence (AI)-assisted quantitative measurement in evaluation of the dynamic changes of CT for COVID-19 pneumonia.Methods:The clinical and chest CT dynamic imaging data of 99 patients with confirmed COVID-19 pneumonia who were hospitalized in Wuhan Central Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January 15, 2020 to March 10, 2020 were retrospectively analyzed. According to the definitive diagnosis, the 99 patients were classified into common ( n=36), severe ( n=33) and critical ( n=30) type, the CT imaging findings of each type were analyzed, including CT basic signs, total volume of pneumonia lesions and percentage of pneumonia lesions of the total lung volume (volume ratio). AI software was used to quantitatively evaluate the dynamic changes of chest CT images. The quantitative indicators included CT peak time of lesions, total volume of lesions peak, volume ratio of lesions peak, maximum growth rate of total volume and maximum growth rate of volume ratio. Kruskal-Wallis rank sum test was used to compare the difference of quantitative indexes between the 3 types, and χ 2 test or Fisher exact probability test was used to compare the difference of qualitative indexes between the 3 types. Sequence measurement and scatter plots were used to show the evolution trend of the volume ratio of the three types of COVID-19 pneumonia lesions. The ROC curve was used to analyze the value of the volume ratio of pneumonia lesions and its maximum growth rate in predicting the conversion of common pneumonia to severe or critical pneumonia. Results:There were statistically significant differences in age and gender distribution among patients with common, severe and critical COVID-19 ( P<0.05), the age of severe and critical types were significantly higher than that of common type ( P<0.01). Compared with common [2.5 (1.0, 5.0) d] and critical type[2.5 (1.0, 4.0) d], the time from onset to the first chest CT scan of severe type was prolonged [5.0 (2.5, 8.0) d, P<0.01]. There were statistically significant differences in involvement of multiple lung lobes (20 cases, 29 cases, 25 cases, χ2=10.403, P=0.006) in patients with common, severe and critical COVID-19 at the first scan, the incidence of the involvement of multiple lung lobes in severe and critical types was significantly higher than that of common type ( P=0.002). The volume ratios of patients with common, severe and critical COVID-19 at the first scan were statistically significant [1.0% (0.2%, 4.7%), 9.30% (1.63%, 26.83%), 2.10% (0.64%, 8.61%), Z=14.236, P=0.001], and the volume ratio of severe type was significantly higher than that of common type ( P<0.001), there was no statistically significant difference between common type and critical type ( P=0.062). Follow-up CT showed that the pneumonia lesions showed a dynamic transformation of progress and recovery, and it was seen that the coexistence of multiphase lesions. The trend line in the scatter plot of the three types of COVID-19 pneumonia lesions showed that the lesions in the advanced stage developed from less to more. The lesion peak volume ratios of the common, severe and critical types were 9.75% (4.83%, 13.18%), 29.80% (23.99%, 42.36%) and 61.81% (43.73%, 72.82%), respectively, the difference was statistically significant ( Z=74.147, P<0.001). The maximum growth rates of lesion volume ratio were 1.27% (0.50%, 1.81%)/d, 4.39% (3.16%, 5.54%)/d and 6.02% (4.77%, 9.96%)/d, respectively, the difference was statistically significant ( Z=52.453, P<0.001). The peak times of lesions were 12.0 (9.0, 15.0) d, 13.0 (10.0, 16.0) d and 16.5 (12.0, 25.0)d, respectively, the difference was statistically significant ( Z=9.524, P=0.009). Taking the volume ratio of pneumonia lesion 22.60% and the maximum growth rate of the volume ratio 1.875%/d as the boundary value, the sensitivity of diagnosing common type to severe or critical type was 92.10% and 96.83%, and the specificity was 100% and 80.56%, respectively. The area under the curve was 0.987 and 0.925, respectively. Conclusions:The lesions of COVID-19 pneumonia show a similar parabolic change on CT imaging. The use of AI technology to dynamitcally and accurately measure the CT pneumonia lesion volume ratio is helpful to evaluate the severity of the disease and predict the development trend of the disease. Patients with a rapid growth of volume ratio are more likely to become severe or critical type.

Mechano-growth factor (MGF) is one of IGF-1 isoforms. MGF is mechanosensitive and has important functions in muscle hypertrophy, regeneration and nerve injury recovery. In this study, MGF cDNA (330 bp) was cloned from stretched osteoblasts by RT-PCR. In order to avoid prolin residue inhibiting enterokinase cleavage, 9bp of MGF cDNA 5' end sequence was truncated by primer, then the obtained truncated MGF (des(1-3)MGF) cDNA (321 bp) was subcloned in pET32a(+) vector to construct a prokaryotic recombination expression plasmid. Trx/des(1-3)MGF fusion protein, existing in forms of solution, was expressed in transformed Escherichia coli strain BL21(DE3) by IPTG induction at 30 degres C. The supernatant of cell lysates was subjected to ion exchange chromatography and Ni2+ metal affinity chromatography, and the fusion protein was obtained with the purity over 95%. After the fusion protein was cleaved by enterokinase, Trx and des(1-3)MGF was isolated by reverse-phase HPLC. Through these procedures, des(1-3) MGF was obtained with the purity of 98%. The protein molecular mass was conformity to the theoretical value by SDS-PAGE and mass spectrometry analysis. The purified des(1-3)MGF was incubated with MC3T3-E1 for cell proliferation and migration assays. The results show that des(1-3)MGF exhibited more facilitative effects on proliferation and migration of MC3T3-E1 than that of des(1-3)IGF-1.
Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Insulin-Like Growth Factor I ; Osteoblasts ; metabolism ; Protein Isoforms ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; STAT5 Transcription Factor ; biosynthesis ; genetics ; Tumor Suppressor Proteins ; biosynthesis ; genetics

This experiment was designed to study the apoptosis and related mechanism of adherent liver tumor cells (SMMC-7721) and adherent normal liver cells (HL-7702) when they were exposed to the steep pulse generated by the steep pulse apparatus for tumor treatment. The results showed that the steep pulse of 200 V could induce tumor cells apoptosis. The tumor cells presented with their apoptosis when they were exposed to the steep pulse from 200 V to 250 V. Laser scanning confocal microscopy was used to make a real time study of calcium burst when the adherent tumor cells were exposed to the steep pulse. The results showed:On the condition of no extracellular Ca2+, the concentration of Ca2+ in tumor cells exposed to the steep pulse of 150 V did not change; the concentration of Ca2+ in tumor cells exposed to the steep pulse of 200 V decreased; the concentration of Ca2+ in tumor cells exposed to the steep pulse of 250 V decreased more evidently. On the condition of existing extracellular Ca2+, the concentration of Ca2+ in tumor cells exposed to the steep pulse of 150 V did not change; the concentration of Ca2+ in tumor cells exposed to the steep pulse of 200 V decreased little; the concentration of Ca2+ in tumor cells exposed to the steep pulse of 250 V reduced little, too. Maybe the change of calcium burst in the tumor cells is the mechanism of apoptosis when cells are exposed to the steep pulse.
Apoptosis ; radiation effects ; Calcium ; metabolism ; Electricity ; Electromagnetic Fields ; Hepatocytes ; cytology ; pathology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Microscopy, Confocal ; Tumor Cells, Cultured

BACKGROUND:To reduce the amount of bleeding and the amount of hemoglobin after total knee replacement has been a key project in the clinical research in the division of bone and joint. Currently, ice therapy has been widely used in the clinic for tissue sweling and pain due to various physical and chemical factors. OBJECTIVE:To investigate the risk factors of postoperative hemoglobin after total knee replacement and discuss the effects of ice intervention. METHODS: 240 patients with osteoarthritis based on the random draw principles were equaly divided into the treatment group and the control group. The general information, disease status, diagnosis and treatment and prognosis of the two groups were investigated. Al patients were actively subjected to artificial total knee replacement. On the basis of the treatment in the control group, the treatment group received ice intervention at 2 hours after replacement for 7 consecutive days. RESULTS AND CONCLUSION:The postoperative hemoglobin decrease occurred in 34 patients, with the incidence of 14.2% among 240 patients at 7 days after replacement. Multivariate logistic regression analysis results showed that age, no ice treatment, body mass index were the main risk factors for hemoglobin decrease after total knee replacement (P < 0.05). Compared with the control group, the postoperative hemoglobin values of the treatment group were significantly higher (P < 0.05). Hemoglobin decrease values, total blood loss, blood transfusion rate, blood transfusion amount, and pain score at 3 and 7 days after replacement were significantly lower in the treatment group than in the control group (P < 0.05). The knee function excelent and good rate was 96.7% in the treatment group, and 95.8% in the control group, which showed no significant difference (P > 0.05). Results verify that clinical application of total knee replacement facilitated the knee recovery in patients with osteoarthritis, but hemoglobin decrease and bleeding existed. Active ice intervention can reduce the risk and relieve postoperative pain.