Objective To observe the eotaxin expression of rat airway smooth muscle cells ( ASMCs) induced by serum from asthmatic rats, and explore the possible mechanism. Methods ASMCs isolated fromrat tracheas were cultured in vivo. Then they were treated with serum from asthmatic rats, or treated with serum and dexamethasone simultaneously. The level of eotaxin protein in supernatant and eotaxin mRNA in ASMCs were measured by ELISA and reverse transcription-polymerase chain reaction. The expression of cAMP in ASMCs was examined by radioimmunoassay. Results After the treatment with sensitized serum, the eotaxin level in supernatant and mRNA expression in ASMCs were significantly higher [ ( 107. 09 ±7. 12) ng/L vs. ( 0. 63 ±0. 56) ng/L, P lt; 0. 05; 1. 39 ±0. 04 vs. 0. 05 ±0. 01, P lt;0. 05] , and the level of cAMP in ASMCs was significantly lower compared with the control group [ ( 17. 58 ±3. 62) ng/L vs. ( 32. 39 ±3. 36) ng/L, P lt; 0. 05] . After intervened by the sensitized serum and dexamethasone simultaneously, the protein and mRNA expressions of eotaxin were lower compared with those intervened by sensitized serumalone [ ( 64. 18 ±4. 04) ng/L and 0. 77 ±0. 19] . The level of eotaxin in supernatant was negatively correlated with cAMP level in ASMCs ( r = - 0. 788, P lt; 0. 01) . Conclusions There is anautocrine function in ASMCs as inflammatory cells after stimulation with sensitized serum. Eotaxin may play an important roll in the pathogenesis of asthma via a cAMP-dependent pathway.
Objective To investigate the delay of the denervated skeletal muscle atrophy with the method of restraining the increment of the connective tissues by tetrandrine and hormone. Methods The left hind limbs of 42 male adult SD rats were made into models of the denervated gastrocnemius, and then the rats were randomly divided into 3 groups, with 14 rats in each. In Group A, tetrandrine (8 mg/L）was injected into the denervated gastrocnemius; in Group B, triamcinolone acetonide（1.6 g/L） was injected; in Group C (the control group),normal saline was injected. Enough samples were obtained according to the different observation indexes at 30 days after operation. Electromyography, muscle wet weight measurement, light microscopy,electron microscopy,and microimage analysis were performed. ResultsThe fibrillation potential amplitude was 0.195 8±0.041 9 μV in Group A and 0.185 2±0.050 3 μV in Group B, and there was no significant difference betweenthe two groups (Pgt;0.05). However,in Group C the fibrillation potential amplitude was 0.137 7±0.058 9μV. The fibrillation potential amplitude was significantly greater in Group A than in Group C（Plt;0.05）. The muscle wet weight was 1.740 0±0.415 9 g in Group A and 1.940 1±0.389 4 gin Group B, and there was no significant difference between the two groups(Pgt;0.05).However, in Group C the muscle wet weight was 0.800 0±0.100 0 g. The muscle wet weight was significantly greater in Group A than in Group C（Plt;0.05）.The microscopy showed that more remarkable atrophy occurred in the control group. The muscle fibers were more complete, thicker and larger, with more nuclei and clearer cross-lines. More connective tissue and flat cells could be observed in Groups A and B. The myogenic protein amount was 440.124 2±46.135 6 in Group A and 476.211 4±41.668 8in Group B, and there was no significant difference between the two groups(Pgt;0.05).However, in Group C the amount was 380.040 0±86.315 9.The myogenic protein amount was significantly greater in Group A thanin Group C（Plt;0.05）. The muscle fiber number, diameter, cross section, and connective tissue increment were all significantly greater in Group A than in Group C（Plt;0.05）; however, there wasno significant difference between Groups A and B (Pgt;0.05). The electron microscopy showed that there were more degeneration changes, such as muscle silk disorder, chondriosome disappearance, and hepatin reduction, could be observed inGroup C than in Groups A and B. Conclusion Tetrandrine and hormone can delay the denervated skeletal muscle atrophy by restraining the increment of the connective tissues.
ObjectiveTo explore the protective effect of low-molecular-weight heparin calcium (LHC) combined with trimetazidine on intestinal smooth muscle of intestinal acute mesangial vein thrombosis (AMVT) in rats and it's mechanism of effect.
MethodsA total of 120 SD male rats were randomly divided into three groups, with 40 rats in each group. LHC group: after the AMVT model established, rats were subcutaneous injection the LHC (30 U/100 g) per 12 h until 72 h after surgery. LHC+trimetazidine group (LHCT group): after the AMVT model established, rats were subcutaneous injection the LHC (30 U/100 g) and tail vein injection the trimetazidine (10 mg/kg) per 12 h until 72 h after surgery. Normal saline group (NS group): after the AMVT model established, rats were subcutaneous injection the NS (0.2 mL/100 g) per 12 h until 72 after surgery. The AMVT model were established by blocking superior mesenteric vein of 8 cm and the edge vein arch. Vena cava blood samples and intestinal segments were collected sequentially at 6 h, 12 h, 24 h, 48 h and 72 h afrer surgery. The levels of malondialdehyde (MDA) and creatine kinase (CK) in the blood, and the level of ATP in the intestinal tissue samples were measured with ELISA. Intestinal tissue were taken from the rats for inestinal tissue section, stained with hematoxylin and eosin, examined under light microscopy and evaluated histopathologically using mesemeche scoring system at different time.
ResultsCompared with the LHC group and NS group, the levels of MDA and CK in blood and histopathology score of intestinal tissues in rats were significantly decreased, and the level of ATP significantly increased in LHCT group at different time point (P < 0.05).
ConclusionTrimetazidine can improve intestinal smooth muscle energy metabolism in the AMVT disease, comined with LHC early can avoid intestinal smooth muscle wall permeability coagulation necrosis and reduce the intestinal smooth muscle damage.
A new method of transfer of the nasolabial skin flap, the myocutaneous flap with pediculated guadratus labii superioris muscle was introduced. It was applied in 9 cases mid-face defects with satisfactory results. The applied anatomy and the its operative technique were briefly discussed.
wenty-one cases with injurys of upper trunk of brachial plexus in 18 and poliomyelitis in 3were treated by transfer of flexor carpi ulnaris muscle to restore flexion of elbow from may, 1981through November, 1992. There were 16 males and 5 females with an average age of 28 years old(ranged 17-60 years). All of the patients was combined with incompetence of abduction function ofshoulder, 6 cases with incompotence of extenxor function of elbow and 11 cases with incompotence ofsupifiation fu...
The noses of eight patients being dead for 2hours were dissected to investigate the layers andstructure of the nose, and the stability of theimplanted silicone noae prosthesis was tested.According to the structure and microstructure ofthe nose studied by us, we suggested a newconcept of nasal muscle and dorsal deepfasciacomplex. We confirmed the prcathesis should beimplanted in the space between the nasal boneand the complex. The reason for complicationhappened in this approach was that...
In order to accurately capture the respiratory muscle movement and extract the synchronization signals corresponding to the breathing phases, a comprehensive signal sensing system for sensing the movement of the respiratory muscle was developed with applying the thin-film varistor FSR402 IMS-C07A in this paper. The system integrated a sensor, a signal processing circuit, and an application program to collect, amplify and denoise electronic signals. Based on the respiratory muscle movement sensor and a STM32F107 development board, an experimental platform was designed to conduct experiments. The respiratory muscle movement data and respiratory airflow data were collected from 3 healthy adults for comparative analysis. In this paper, the results demonstrated that the method for determining respiratory phase based on the sensing the respiratory muscle movement exhibited strong real-time performance. Compared to traditional airflow-based respiratory phase detection, the proposed method showed a lead times ranging from 33 to 210 ms [(88.3 ± 47.9) ms] for expiration switched into inspiration and 17 to 222 ms [(92.9 ± 63.8) ms] for inspiration switched into expiration, respectively. When this system is applied to trigger the output of the ventilator, it will effectively improve the patient-ventilator synchrony and facilitate the ventilation treatment for patients with respiratory diseases.
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.
Objective To analysis the electrophysiological dominance weight of the triceps brachii muscle/extensordigitorum communis muscle innervated by brachial plexus and to conclude its effect on the ipsilateral C7 transfer so as to offer electrophysiological data for the safety and indication of i psilateral C7 transfer. Methods From August 2007 to October 2007, 15 patients with complete brachial plexus nerve root avulsion received contralateral C7 transfer. There were 13 males and 2 females aged 18-49 years (28 years on average). Injury was caused by fall ing in 1 case, by crush in 2 cases and by traffic accident in 12 cases, involving left side in 8 cases and right side in 7 cases. The upper, middle and lower trunk of the brachial plexus were stimulated respectively, the compound muscle action potential (CMAP) at the triceps brachii muscle/extensor digitorum communis muscle was recorded, and then the electrophysiological dominance weight of the triceps brachii muscle/extensor digitorum communis muscle innervated by brachial plexus was confirmed according to the comparison of the ampl itude percentage of the CMAP by three trunks. The muscle strength of triceps brachii muscle/extensor digitorum communis muscle was evaluated and the electromyogram was taken 6 months after operation. Results All patients were followed up for 6 months. Concerning the electrophysiological dominance weight, the triceps brachii muscle was mainly innervated by uppermiddle trunk in 3 cases (20%), by middle-lower trunk in 3 cases (20%), by whole trunk in 7 cases (47%) and by middle trunk in 2 cases (13%). While the extensor digitorum communis muscle was mainly innervated by middle-lower trunk in 3 cases (20%), by whole trunk in 10 cases (67%) and by lower trunk in 2 cases (13%). Concerning the triceps brachii muscle, 2 patients got the muscle strength of 4 grade with recruitment simple phase at 1 month after operation and returned to normal at 3 month after operation, while 13 patients got the muscle strength of 5 grade with recruitment simple or mixed phase at 1 month after operation. Concerning the extensor digitorum communis muscle, the muscle strength and the recruitment phase of all 15 patients recovered to normal at 1 month after operation. Conclusion To patients with various kinds of electrophysiological dominance weight, the cutting of C7 does not substantially damage the triceps brachii muscle or extensor digitorum communis muscle, indicating that the ipsilateral C7 transfer is safe and feasible. However, it should be appl ied prudently for the patients with high dominance weight since it may result in the short-term decrease of triceps brachii muscle strength.
Objective To investigate the effect of various concentration of platelet-rich plasma (PRP) on osteogenic differentiation of rabbit skeletal muscle-derived stem cells (SMSCs) cultured in vitro. Methods Blood drawn from the central ear arteries of 9 one-year-old New Zealand white rabbits weighing 2.5-3.0 kg (male and female) was used to prepare PRP (Landesberg method). Full blood count and platelet count in PRP were tested. Soleus muscle of right hindl imb in rabbit was obtained and used to culture SMSCs in vitro. The cells at passage 3 were randomly divided into different groups: the experimental groups in which the cells were treated by conditioned culture media with various concentrations of autologousPRP (6.25%, 12.50%, 25.00%, 50.00%), and the control group in which the cells were treated with the media without PRP. At different time points after intervention, osteogenetic activity of the cells was detected by ALP staining observation, ALP activity detection was conducted, al izarin red staining for calcium nodules and immunofluorescence staining for osteocalcin were performed, and core binding factor α1 (Cbfα1) of osteogenic gene expression was tested by RT-PCR. Results The full blood PRP count and the platelet count in PRP was (3.06 ± 0.46) × 105/μL and (18.08 ± 2.10) × 105/μL, respectively. ALP staining: the cells in all the experimental groups were positive for the staining with many black sediment particles in cytoplasm; the cells in the control group were negative staining. ALP activity: all the experimental groups were higher than the control group (P lt; 0.05), the experimental group at 12.50% was superior to other experimental groups at each time point (P lt; 0.05). Al izarin red staining: at 14 days after culture, orange-red calcium nodules were evident in all the experimental groups; no orange-red calcium nodules were observed in the control group with a mineral ization rate of zero; there were significant difference between the experimental groups and the control group in terms of mineral ization rate (P lt; 0.05), the experimental group at 12.50% had a higher mineral ization rate than other experimental groups (P lt; 0.05). Immunofluorescence staining for osteocalcin: at 7 days after culture, the experimental groups were positive for the staining with yellow fluorescence in cytoplasm, and the result of the control group was negative. RT-PCR detection: no obvious changes of the gene expression were noted at 4, 12, and 24 hoursafter culture in the control group; the gene expression in all the experimental groups was significant superior to that of control group, especially at 12 hours, and the expression in the experimental group at 12.50% was the highest. Conclusion PRP can obviously promote the osteogenic differentiation of SMSCs cultured in vitro in a concentration-dependent manner, and the 12.50% is proved to be the ideal concentration.
