Objective To explore the in vitrodifferentiation of the rat mesenchymal stem cells (MSCs ) into the skeletal muscle cells induced by the myoblast differentiation factor (MyoD) and 5-azacytidine. Methods The MSCs were taken from the rat bone marrow and the suspension of MSCs was made and cultured in the homeothermia incubator which contained 5% CO2at 37℃. The cells were observed under the inverted phase contrast microscope daily. The cells spreading all the bottom of the culture bottle were defined as onepassage. The differentiation of the 3rd passage of MSCs was induced by the combination of 5-azacytidine, MyoD, transforming growth factor β1, and the insulin like growth factor 1. Nine days after the induction, the induced MSCs were collected, which were analyzed with the MTT chromatometry, theflow cytometry, and the immunohistochemistry. Results The primarily cultured MSCs grew as a colony on the walls of the culture bottle; after the culture for 5-7 days, the cells were shaped like the fibroblasts, the big flat polygonal cells, the medium sized polygonal cells, and the small triangle cells; after the culture for 12 days, the cells were found to be fused, spreadingall over the bottle bottom, but MSCs were unchanged too much in shape. After the induction by 5-azacytidine, some of the cells died, and the cells grew slowly. However, after the culture for 7 days, the cells grew remarkably, the cell volume increased gradually in a form of ellipse, fusiform or irregularity. After theculture for 14 days, the proliferated fusiform cells began to increase in a great amount. After the culture for 18-22 days, the myotubes increased in number and volume, with the nucleus increased in number, and the newly formed myotubes and the fusiform myoblst grew parallelly and separately. The immunohistochemistry for MSCs revealed that CD44 was positive in reaction, with the cytoplasm ina form of brown granules. And the nucleus had an obvious border，and CD34 was negative. The induced MSCs were found to be positive for desmin and specific myoglobulin of the skeletal muscle. The flow cytometry showed that most of the MSCs and the induced MSCs were in the stages of G0/G1，accounting for 79.4％ and 62.9％，respectively; however, the cells in the stages of G2/S accounted for 20.6％ and 36.1％. The growth curve was drawn based on MTT，which showed that MSCs weregreater in the growth speed than the induced MSCs. The two kinds of cells did not reach the platform stage，having a tendency to continuously proliferate.ConclusionIn vitro,the rat MSCs can be differentiated into the skeletal muscle cells with an induction by MyoD and 5-azacytidine, with a positive reaction for the desmin and the myoglobulin of the skeletal muscle. After the induction, the proliferation stage of MSCs can be increased, with a higher degree of the differentiation into the skeletal muscle.
ObjectiveTo evaluate the superiority of nasopharyngeal airway on obesity patients during general anesthesia induction period. MethodForty-two trachea cannula and general anesthesia obesity patients treated from June to November in 2013 were chosen and divided equally into two groups:nasopharyngeal airway group (group A) and control group (group B). Mean arterial pressure (MAP), heart rate (HR), pulse oxygen saturation (SpO2), arterial blood partial pressure of carbon dioxide (PaCO2) were recorded when the patients entered the operation room, three minutes after man-made positive pressure ventilating and five minutes after intubation. Peak voltage (Ppeak) of man-made positive pressure ventilation for three minutes was also observed, and intubation frequency and time, mouth mucosa bleeding, and sore throat examples were compared between the two groups. ResultsCompared with group B, MAP, HR, PaCO2 and Ppeak three minutes after man-made positive pressure ventilating were lower (P<0.05), but SpO2 was higher in group A (P<0.05). Intubation frequency and time, mouth mucosa bleeding, and sore throat examples of group A were less than those in group B (P<0.05). ConclusionsNasopharyngeal airway is better for obesity patients during general anesthesia induction period, which also improves anesthesia safety level.
In order to explore further the regulatory factors to the potentiality in inducing osteogenesis by fibroblasts, the fibroblasts were isolated, and purified from human skin, and were grown in incubation in the media of EGF, IL-6, TNF-alpha and BMP2 at different concentrations for two weeks, then, the markers for osteogenic features were investigated by biochemistry, histochemistry and electron microscopic observations. It was found that the combined use of TNF-alpha and BMP2 could stimulate fibroblasts to secrete alkaline phosphatase, osteocalcin and collagen, and the morphological changes of the fibroblasts were also very striking. In the extracellular matrix, the collagen fibrils, with or without periodicity, were arranged regularly or randomly oriented, and numerous minute calcium granules were interspersed among them. The fibroblasts were interwoven one on top of another in the form of multilayer structure and on the surface, there were secreting granules and piling up of calcium crystals which coalessed steadily and increased in size in forming bony nodules. It was considered that TNF-alpha and BMP2 were capable of inducing the fibroblasts to form bone.
Objective To explore the regulator factor of osteogenes is induced by the fibroblast in vitro so as to provide enough seeding cells for the bon e tissue engineering. Methods The fibroblasts were isolated and purified from granu lation of New Zealand rabbits, and they were incubated in the media offibronectin (FN) 10, 20, 40, 60 and 80 μg/ml, respectively, in the experimenta l grou ps 1- 5，but there was no FN in the control group. The markers for osteogenic features were investigated by fibroblast morphogenesis,calcium nodules formationratios,labeling of tetracycline fluorescence, labeling of 3H-TdR, determination of o steocaline, and labeling of 3H-proline within 2 weeks. Results The morphologic al changes of the fibroblasts were manifested as transference from a long spindle to a round or multiple form, shifted nucleus increased in number, confluenced and formed multilayered structure. There was a piling-up of calcium crystals that were gradually merged into foggy substances. The foggy substances increased and formed nodules. The calcium nodules formation ratios were as follows: 15.35％± 3.45％in the control group, and 53.73％± 9.49％, 75.21％± 9.80％, 98.34％± 15.2 0％, 61.83％± 10.04％, and 45.11％± 8.70％ in the experimental groups 1.5 ,respectively. There was a significant difference between the control group and the 5 experimental groups at 14 days (Plt;0.05), and a significant differenc e be tween the experimental group 3 and the other experimental groups at 14 days (Plt;0.05). The histochemical study on the nodules with the specific labeling of tet racycline fluorescence indicated that the nodules were composed of new bones. Conclusion Fibronectin can stimulate the fibroblast to prolifer ate, secrete osteocaline, and synthesize collagen fibrils. Fibronectin, in an optimal dose of 40 -60 μg/ml, is capable of inducing the fibroblast to form the bone.
ObjectiveTo investigate the feasibility of adipose-derived mesenchymal stem cells (ADMSCs) differentiating into corneal epithelium-like cells after transfection with Pax6 gene. MethodsThe adipose tissue from bilateral inguinal of healthy C57BL/6 mice (5-6 weeks old) was used to isolate and culture ADMSCs.The 3rd passage ADMSCs were subjected to treatments of non-transfection (group A),pcDNA3.1 empty vector transfection (group B),and recombinant plasmid of pcDNA3.1-Pax6 transfection (group C),respectively.At 48 hours after transfection,the cells in groups B and C were selected with G418.The cell morphology changes were observed under the inverted microscope.Pax6 protein and level of corneal epithelial cells specific molecular-cytokeratin 12 (CK-12) were measured by Western blot.Real-time fluorescence quantitative PCR was applied to measure the mRNA expression of CK-12. ResultsNo morphology change was observed in groups A and B.Two different cell clones were found in group C.No.1 selected clone showed a flagstone-like appearance that was similar to that of corneal epithelial cells;No.2 selected clone showed a net-like appearance,with 3-7 cell processes.The Western blot results showed the Pax6 protein expression in 2 clones of group C,but no expression in groups A and B; and CK-12 protein expression was only observed in No.1 selected clone of group C,and no expression in the others.The real-time fluorescence quantitative PCR results showed that the CK-12 mRNA expression level of No.1 selected clone of group C was 8.64±0.73,which was significantly higher than that of No.2 selected clone of group C (0.55±0.42),group B (1.36±0.40),and group A (1.00±0.00) (P<0.05),and there was no significant difference among groups A,B and No.2 selected clone of group C (P>0.05). ConclusionPax6 gene transfection could induce differentiation of ADMSCs into corneal epithelium-like cells which express CK-12 at both the mRNA and protein levels.This result provides a promising strategy of generating corneal epithelilcm-like cells for construction of tissue engineered cornea.
The optimal treatment of stage ⅢA-N2 non-small cell lung cancer （NSCLC） remains controversial. Resultsof primary surgery alone are not satisfied. Surgery after induction chemotherapy yields better outcomes compared to resectiononly which has been widely accepted. Randomized studies show induction chemotherapy followed by either radiotherapy or surgery have approximately equivalent survival outcomes，significant improved survival can be achieved by combined surgery in selected patients. Low-grade N2，effective response and mediastinal downstaging after induction therapy，and successful complete resection by lobectomy，are good indications of surgery. Ideal treatments are approached base on theheterogeneity of N2 . Patients with bulky or fixed N2 disease should be considered for radical chemo-radiotherapy，and surgeryshould be a part of multi-modality management for patients with non-fixed，non-bulky，single-zone N2 disease. Further randomized trials of surgery added to multi-modality management in patients with multi-zone N2 disease should be taken in order to establish possible subgroups of patients might be benefitted more from the addition of surgery.
Objective To review the research progress of the current methods of inducing bone marrow mesenchymal stem cells (BMSCs) to chondrogenic differentiation in vitro so as to provide references for researches in cartilage tissue engineering. Methods Various methods of inducing BMSCs differentiation into the chondrogenic l ineage in vitro inrecent years were extensively reviewed and analyzed. Results Adding exogenous growth factors is still the mainly methodof inducing BMSCs differentiation into the chondrogenic l ineage; among the members, transforming growth factor β (TGF-β) family is recognized as the most important chondrogenic induction factor. Other important inducing factors include various chemical factors, physical factors, transgenic methods, and the microenvironmental induction. But the problems of low inducing efficiency and unstable inducing effects still exist. Conclusion The progress of chondrogenic induction of BMSCs promotes its util ization in cartilage tissue engineering. Further researches are needed for establ ishing more efficient, simpler, and safer inducing methods.
ObjectiveTo study the inducting differentiation effect of the sciatic nerve extracts on rabbit adipose-derived stem cells (ADSCs) in vitro. MethodsThe ADSCs were isolated from 2 healthy 4-month-old New Zealand rabbits (weighing, 2.0-2.5 kg) and cultured to passage 3, which were pretreated with 10 ng/mL basic fibroblast growth factor (bFGF) for 24 hours before induction. Then the induction media containing the extracts of normal sciatic nerve (group B) and injured sciatic nerve at 3, 7, and 14 days (group C, group D, and group E) were used, and D-Hank was used in group A as blank control group. The morphological changes of the cells were observed. At 7 days of induction, the gene expressions of neuron-specific enolase (NSE), nestin (NES), and S-100 were detected by real-time fluorescent quantitative PCR. The S-100 protein expression was tested by immunocytochemical staining. ResultsAt 4 days after induction, some ADSCs of groups C, D, and E showed the morphology of Schwann-like cells or neuron-like cells, the change of group D was more obvious; and the ADSCs of group A and B had no obvious change, which were still spindle. The S-100 immunocytochemical staining showed positive expression in groups C, D, and E (more obvious in group D) and negative expression in groups A and B. The gene expression of S-100 displayed time-dependent increases in groups C and D, which was significantly higher than that of groups A, B, and E (P<0.05), but no significant difference was found between groups C and D (P>0.05). The gene expression of NSE showed the same tendency to S-100, which reached the peak in group D; the gene expression of NSE in groups D and E was significantly higher than that of groups A, B, and C (P<0.05), and groups D and E showed significant difference (P<0.05). However, the gene expression of Nestin showed no significant difference among different groups (P>0.05). ConclusionThe ADSCs can be induced to differentiate into Schwann-like cells or neuron-like cells with sciatic nerve extracts; and the early stage (3-7 days) after injury is the best time for stem cell transplantation.
Because of the diversity and complexity of clinical indicators, it is difficult to establish a comprehensive and reliable prediction model for induction of labor (IOL) outcomes with existing methods. This study aims to analyze the clinical indicators related to IOL and to develop and evaluate a prediction model based on a small-sample of data. The study population consisted of a total of 90 pregnant women who underwent IOL between February 2023 and January 2024 at the Shanghai First Maternity and Infant Healthcare Hospital, and a total of 52 clinical indicators were recorded. Maximal information coefficient (MIC) was used to select features for clinical indicators to reduce the risk of overfitting caused by high-dimensional features. Then, based on the features selected by MIC, the support vector machine (SVM) model based on small samples was compared and analyzed with the fully connected neural network (FCNN) model based on large samples in deep learning, and the receiver operating characteristic (ROC) curve was given. By calculating the MIC score, the final feature dimension was reduced from 55 to 15, and the area under curve (AUC) of the SVM model was improved from 0.872 before feature selection to 0.923. Model comparison results showed that SVM had better prediction performance than FCNN. This study demonstrates that SVM successfully predicted IOL outcomes, and the MIC feature selection effectively improves the model’s generalization ability, making the prediction results more stable. This study provides a reliable method for predicting the outcome of induced labor with potential clinical applications.
Objective To investigate the curve correlation between ventilation pressure and tidal volume in assisted mechanical ventilation with facemask during anesthesia induction. Methods Between January and August 2015, 120 patients, American Society of Anesthesiology Ⅰ-Ⅱ, undergoing selective gynecological surgery were randomly divided into four groups: groups P5, P10, P15 and P20, with 30 patients in each group. Mask ventilation pressure for the four groups were respectively 5, 10, 15 and 20 cm H2O (1 cm H2O=0.098 kPa). Patients were ventilated by preset ventilation pressure and frequency based on different groups after loss of consciousness. Mean ventilation volume (mean value of three tidal volumes) and end-tidal carbon dioxide pressure (PetCO2) were recorded for analysis. Results There was no significant difference among the four groups in patient’s general condition (P>0.05). The tidal volume of assisted mechanical ventilation increased with ventilation pressure degrees, and the differences among the four groups were significant (P<0.05). After curve regression analysis, tidal volume and ventilation pressure showed a positive linear correlation when ventilation pressure was set at 5-20 cm H2O, and the correlation equation was: tidal volume = 33.612×ventilation pressure-53.155. PetCO2 in P5 group was lower than those in the other three groups (P<0.05), while there were no significant differences among groups P10, P15 and P20 (P>0.05). Conclusion When ventilation pressure is set at 5-20 cm H2O in assisted mechanical ventilation with facemask during anesthesia induction, tidal volume and ventilation pressure show a positive linear correlation.