In order to identify the differentially expressing gene of bone marrow mesenchymal stem cells (MSCs) stimulated by electromagnetic field (EMF) with osteogenesis microarray analysis, the bone marrow MSCs of SD rats were isolated and cultured in vitro. The third-passage cells were stimulated by EMFs and total RNA was extracted, purified and then used for the synthesis of cDNA and cRNA. The cRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray respectively. The hybridization signals were acquired by using X-ray film after chemiluminescent detection and the data obtained were analyzed by employing the web-based completely integrated GEArray Expression Analysis Suite. RT-PCR was used to identify the target genes: Bmp1, Bmp7, Egf and Egfr. The results showed that 19 differentially expressing genes were found between the stimulated group and the control group. There were 6 up-regulated genes and 13 down-regulated genes in the stimulated group. Semi-quantitative RT-PCR confirmed that the expressions of Bmp1, Bmp7 mRNA of the stimulated group were up-regulated (P<0.05) and those of Egf, Egfr were down-regulated (P<0.05). It was suggested that the gene expression profiles of osteogenesis of the bone marrow MSCs were changed after EMF treatment. It is concluded that the genes are involved in skeletal development, bone mineral metabolism, cell growth and differentiation, cell adhesion etc.
Bone Marrow Cells/*cytology ; Cell Differentiation ; Down-Regulation ; Electromagnetic Fields ; Gene Expression Profiling ; Gene Expression Regulation ; Mesenchymal Stem Cells/*cytology ; Nucleic Acid Hybridization ; Oligonucleotide Probes/chemistry ; Osteogenesis/*genetics ; RNA, Complementary/metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To study the influence of pulsed electromagnetic fields on the expression of type I collagen by bone marrow mesenchymal stem cells and it's mechanism. Methods The bone marrow mesenchymal stem cells of SD rats were isolated and cultured in vitro.The third passage cells were harvested and exposed to pulsed electromagnetic fields(PEMFs)at 15 Hz and 1 mT 8 h/d for 3 days.A semi-quantitative RT-PCR technique was used to measure the type I collagen mRNA expression;ELISA and immunohistochemitistry techniques were used to measure type I collagen expression.Inhibitors and promoters of the cAMP-dependent protein kinase A(cAMP-PKA)pathway were added.After the cAMP-PKA pathway had been inhibited or promoted,the effects of the PEMF on type I collagen expression were measured again using ELISA and immunohistoehemistry.Results PEMFs at 15 Hz and 1 mT induced significant promotion of the expression of type I collagen(P≤0.01)in comparison with the controls. The type I collagen expression was reduced when the cAMP-PKA pathway inhibitor H-89 was added,and raised when the promoter 8-Br-cAMP was added.Conclusion PEMFs at 15 Hz 1 mT can promote type I collagen expression of bone marrow mesenchymal stem cells.and the effect is correlated with the cAMP-PKA pathway.

Objective To investigate the effects of an electromagnetic field on the extra-cellularly regulated kinase(ERK)signalling pathway and to determine the impact of electromagnetic activation on osteogenic proliferation and differentiation in rat bone marrow mesenchymal stem cells.Methods Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro.The third-passage cells were divided into 4 groups(Control,PD98059,EMF and EMF+PD98059).Western blotting Was used to detect the activation of the ERK signal pathway after exposure to an electromagnetic field.MTT assay Was used to determine the activation of proliferation in the celb in the different groups.The cells' alkaline phosphatase activities were also detected. Results (1)The ERK signal pathway in these rat bone marrow mesenchymal stem cells was activated after exposure to a 15 Hz.1 mT,sine wave form electromagnetic field for 5 min.Activation remained high for at least 1 h.PD98059 can effectively block the activation of the ERK signal pathway.(2)Cell proliferation was promoted after exposure to the electromagnetic field,and this effect could be significantly inhibited by PD98059.(3)Alkaline phosphatase was significantly elevated in these bone marrow mesenchymal stem cells after exposure to the electromagnetic field.The activation in the EMF+PD98059 group Was slightly greater than in the EMF group.Conclusion Electromagnetic fields of 15 Hz and 1 mT can activate the ERK signal pathway and alter proliferation and osteogenic differentiation in the bone marrow mesenchymal stem cells of rats.

In order to identify the differentially expressing gene of bone marrow mesenchymal stem cells (MSCs) stimulated by electromagnetic field (EMF) with osteogenesis microarray analysis, the bone marrow MSCs of SD rats were isolated and cultured in vitro. The third-passage cells were stimulated by EMFs and total RNA was extracted, purified and then used for the synthesis of cDNA and cRNA. The cRNA of stimulated group and the control group was hybridized with the rat oligo osteogenesis microarray respectively. The hybridization signals were acquired by using X-my film after chemiluminescent detection and the data obtained were analyzed by employing the web-based completely integrated GEArray Expression Analysis Suite. RT-PCR was used to identify the target genes: Bmpl, BmpT, Egf and Egfr. The results showed that 19 differentially expressing genes were found between the stimulated group and the control group. There were 6 up-regulated genes and 13 down-regulated genes in the stimulated group. Semi-quantitative RT-PCR confirmed that the expres- sions of Bmpl, Bmp7 mRNA of the stimulated group were up-regulated (P<0.05) and those of Egf, Egfr were down-regulated (P<0.05). It was suggested that the gene expression profiles of osteogene- sis of the bone marrow MSCs were changed after EMF treatment. It is concluded that the genes are involved in skeletal development, bone mineral metabolism, cell growth and differentiation, cell ad- hesion etc.

Objective: To evaluate the clinical value of three-dimensional ultrasound VOCAL technique in evaluating gastric emptying function in children with functional dyspepsia (FD). Methods: Seventy-one children with FD who were treated in Wenzhou Integrated Traditional Chinese and Western Medicine Hospital Affiliated to Zhejiang University of Traditional Chinese Medicine from January 2018 to June 2018 were enrolled as the study subjects (observation group), and 71 normal children without FD were selected as controls (control group). The gastric emptying, antral pyloric systolic contraction frequency and distal gastric contraction movement in different time groups were analyzed, and GET1/2 and 2h gastric residual situation before and after treatment in the observation group were compared. Results: The GET1/2 of the observation group was (60.2±12.69)min, and the gastric emptying rate of the observation group was (61.9±12.2)min and (72.0±12.3)min at 90min and 120min, which were significantly better than those of the control group, and the differences were statistically significant (t=19.092, 15.092, 14.882, P=0.016, 0.024, 0.033). The frequency of antral pyloric systolic contraction, gastric antrum contraction frequency and gastric antrum contraction amplitude were (1.1±0.7)times/min, (2.9±0.8)times/min, (1.0±0.6)cm, respectively.Compared with the control group, the differences were statistically significant (t=16.092, 21.224, 8.092, all P<0.05). In the observation group of 71 children: GET1/2 before treatment and after treatment were (68.1±11.8)min, (54.2±9.8)min, respectively.The gastric residual at 2h after treatment was (31.2±8.0)%, (22.1±7.0)%, respectively.The GET1/2 and 2h gastric residual rate of the children were significantly lower than before treatment, and the differences were statistically significant (t=18.983, 21.004, all P<0.05). Conclusion Three-dimensional ultrasound VOCAL technology can be used as an evaluation standard for FD in children.

Objective:To investigate the effects of airway humidification therapy on pertussis in infants.Methods:Sixty children with pertussis who received treatment in Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine from May 2020 to September 2021 were included in this study. They were randomly divided into study and control groups, with 30 infants per group. The control group was treated with conventional drugs. The study group was subjected to airway humidification therapy based on conventional drug treatment. Clinical efficacy was evaluated in each group. Time to an improvement in clinical symptoms and length of hospital stay were recorded. Lung function indicators and serum inflammatory factor levels were determined before and after treatment. Pertussis-related complications were recorded.Results:The overall response rate in the study group was significantly higher than that in the control group (100.00% vs. 76.7%, χ2 = 5.82, P = 0.016). Time to disappearance of spastic cough, time to resolution of cyanosis, time to disappearance of pulmonary rales, and length of hospital stay in the study group were (3.10 ± 0.67) days, (1.53 ± 0.68) days, (4.27 ± 0.58) days, and (11.57 ± 0.73) days, respectively, which were significantly shorter than (4.23 ± 1.99) days, (2.67 ± 1.77) days, (5.63 ± 2.38) days, (13.33 ± 2.40) days ( t = 2.93, 3.27, 3.04, 3.86; P = 0.005, 0.002, 0.003, P < 0.001). Respiratory rate in the study group was significantly lower than that in the control group [(21.83 ± 1.15) breaths/minute vs. (24.23 ± 3.12) breaths/minute, t = 3.94, P = 0.002]. Tidal volume/kg body weight, the ratio of time to peak expiratory flow to total expiratory time and the ratio of volume to peak expiratory flow to total expiratory volume in the study group were (10.70 ± 0.81) mL/kg, (41.60 ± 1.57)%, (42.89 ± 1.44)%, respectively, which were significantly higher than (9.78 ± 1.34) mL/kg, (39.18 ± 3.37)%, (40.20 ± 3.05)% in the control group ( t = 3.21, 3.56, 4.36; P < 0.001, P = 0.001, P < 0.001). Serum tumor necrosis alpha (TNF-α), interleukin-6 and interleukin-8 levels in the study group were (62.44 ± 2.96) ng/L, (46.59 ± 1.96) ng/L, (54.63 ± 3.27) ng/L, respectively, which were significantly lower than (79.86 ± 3.30) ng/L, (58.20 ± 2.10) ng/L, (63.31 ± 3.86) ng/L in the control group ( t =21.53, 22.13, 9.38, all P < 0.001). The overall incidence of pertussis in the study group was significantly lower than that in the control group (3.3% vs. 26.7%, χ2 = 4.71, P = 0.030). Conclusion:Airway humidification therapy can shorten the time to reduction of symptoms of spasmodic pertussis, improve clinical efficacy, strengthen pulmonary ventilation function, lower respiratory tract inflammatory responses and reduce the risk of pertussis-related complications.

Objective To investigate the clinical,laboratory and imaging characteristics of children with Mycoplasma pneumoniae pneumonia (MPP) in different age groups.Methods One hundred and sixty-four children whose ages were 1 month to 14 years old with MPP hospitalized at Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine in 2017 were enrolled as study objects.All children were classified into 2 groups:1 month to 3 year-old group (46 cases) and ＞ 3 to 14 year-old group (118 cases).The clinical features,main laboratory and imaging findings of 2 groups were retrospectively analyzed and compared.Results (1) Compared with 1 month to 3 year-old group,＞ 3 to 14 year-old group had more patients with fever [97.5％ (115/118 cases) vs.84.8％ (39/46 cases)],longer duration of fever [48.3％ (57/118 cases) vs.23.9％ (11/46 cases)] and higher fever [83.1％ (98/ 118 cases) vs.52.2％ (24/46 cases)],but less patients with wheezing [2.5％ (3/118 cases) vs.43.5％ (20/46 cases)],shortness of breath and dyspnea [0.8％ (1/118 cases) vs.15.2％ (7/46 cases)],dry rale [1.7％ (2/118 cases) vs.60.9％ (28/46 cases)] and wet pulmonary rale [50.0％ (59/118 cases) vs.69.6％ (32/46 cases)],and the differences were statistically significant (all P ＜ 0.05).(2) The increase of platelet count,creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) in 1 month to 3 year-old group of MPP [316.0 (229.3,372.3) × 109/L,25.70 (17.70,31.98) U/L,346.5 (310.3,388.3) U/L] were more common than those in ＞ 3 to 14 year-old group [266.0 (205.8,317.8) × 109/L,21.50 (15.75,28.00) U/L,303.0 (173.0,352.0) U/L],while the increase of C reactive protein (CRP) and erythrocyte sedimentation rate (ESR) in ＞ 3 to 14 year-old group [12.66(9.16,19.44) mg/L,23.00(17.75,29.00) mm/1 h] were more common than those in 1 month to 3 year-old group[2.46 (0.54,11.63) mg/L,14.00 (10.00,20.25) mm/1 h],and there were significant statistical differences (all P ＜ 0.05).(3) The MPP imaging features of children in different age groups were diverse.In 1 month to 3 year-old group,the double lung involvement [80.4％ (37/46 cases)] was more obvious than that in ＞3 to 14 year-old group [55.9％ (66/118 cases)],and the difference was statistically significant (P ＜ 0.05).The lesions of 1 month to 3 year-old group were more common in lobular pulmonary [82.6％ (38/46 cases)],while in ＞ 3 to 14 year-old group,the radiographic findings were predominant with consolidation in the segment or lobe of the lung [33.1％ (39/118 cases)],and the difference was statistically significant (P ＜ 0.01).Conclusions Younger than 3 year-old children with MPP are prone to wheezing,shortness of breath,dyspnea and lung rale,while the rise of CK-MB and LDH is common.Imaging is mainly involved in double lung involvement and pulmonary lobule infiltration.The older the children with MPP are,the longer the fever duration is,and more common is the higher fever.The older the children,the more more easily they have the relative increase of CRP and ESR,and at the same time they are more likely to have the consolidation of the lung segment or lobe.

Hydrogels are hydrophilic three-dimensional networks that exhibit soft and flexible textures after absorbing water, making them biocompatible and biodegradable. Due to their similarity to biological tissues, hydrogels find extensive applications in the field of biomedical mechanism. In this paper, the preparation methods of physical hydrogels and chemical hydrogels are introduced according to the different cross-linking agents, and the application progress of hydrogels in cell therapy is reviewed.