Objective To explore the method that can inducethe mesenchymal stem cells (MSCs) to differentiate into the neuronlike cells in vitro．Methods The neuron-like cells were isolated froman SD rat (age, 3 months; weight, 200 g). They underwent a primary culture; theinduced liquid supernatant was collected, and was identified by the cell immunohistochemistry. The C3H1OT1/2 cells were cultured, as an MSCs model, and they were induced into differentiation by β-mercaptoethanol (Group A) and by the liquid supernatant of the neuron-like primary cells (Group B), respectively. The cells were cultured without any induction were used as a control (Group C). Immunohistochemistrywas used to identify the type of the cells． Results The result of the immunochemistry showed that the cells undergoing the primary culture expressed the neurofilament protein (NF) and the neuronspecific enolase (NSE), and they were neuron-like cells. β-mercaptoethanol could induce the C3H1OT1/2 cells toexpress NF and NSE at 2 h, and the expression intensity increased at 5 h. The liquid supernatant of the primarily-cultured neuron-like cells could induce theC3H1OT1/2 cells to express NF and NSE at 1 d, but the expression intensity induced by the liquid supernatant was weaker than that induced by β-mercaptoethanol. The positivity rate and the intensity expression of NSE were higher than those of NF. Conclusion MSCs can differentiate into the neuron-like cells by β-mercaptoethanol and the microenvironment humoral factor, which can pave the way for a further study of the differentiation of MSCs and the effectof the differentiation on the brain trauma repair.
Objective To investigate the myogenic differentiation of mesenchymal stem cells (MSCs) after being transplanted into the local muscle tissues. Methods The serious muscleinjured model was established by the way of radiation injury, incising, and freezing injury in 36 mouses. Purified MSCs derived from bone marrow of male mouse and MSCs induced by5-azacytidine(5-Aza-CR) were transplanted into the local of normal muscle tissues and injured muscle tissues of femal mouse. The quantity of MSCs and the myogenic differentiation of implanted MSCs were detected by the method of double labeling, which included fluorescence in situ DNA hybridization (FISH) and immuno-histochemistry on the 1st, 3rd, 6th, 9th, 12th, and 15th day after transplantation. Results The quantity of implanted MSCs decreased as timepassed. MSCs’ differentiation into myoblasts and positive expression of desmin were observed on the 15th day in purified MSCs group and on the 6th day in induced MSCs groups. Conclusion MSCs could differentiate into myoblasts after being implanted into the local of muscle tissues. The differentiationoccurs earlier in the induced MSCs group than that in purified MSCs group.
Objective To explore the effects of mechanical stretch with variant frequencies on the alignment and differentiation of the multilayer myotubes cultured in vitro, and to select the optimized cultural condition of regenerative skeletal muscle tissue with stress loading cultured in vitro. Methods C2C12 myoblasts cultured in vitro in the groove casts of Sylgard 184 were induced into the multilayer myotubes. Meanwhile the myoblasts were treated with various mechanical stretch withcells tensile instrument, at the amplitude of 10% and the frequency of 0 (group A), 0.25 (group B), 0.50 (group C), and 1.00 Hz (group D) for 1 hour, 3 times a day. The myotubes morphology was observed by inverted phase contrast microscope at 5, 7, and 10 days after continuous mechanical stretch. And the expressions of mRNA for myogenic differentiation antigen (MyoD), Myogenin, Desmin, and myosin heavy chain (MyHC) were detected by RT-PCR and real-time fluorescent quantitative PCR (QRT-PCR), respectively. Results The mechanical stretch could promote the al igned fusion and increase the number of myotubes. Indeed the multilayer myotubes arranged more closely in group B at 7 days. At the same group, as the time went on, the mRNA expressions of MyoD gradually decl ined in each group. There were significant differences in mRNA expressions of MyoD between 5 days and 7, 10 days (P lt; 0.05). The mRNA expressions of Myogenin, Desmin, and MyHC were highest at 7 days. There were significant differences between different time points (P lt; 0.05), except the mRNA expression of Desmin of group B between 7 and 10 days (P gt; 0.05). At the same time, with the increase of frequency, the highest mRNA expressions of MyoD, Myogenin, Desmin, and MyHC were in group B. There were significant differences at the same time between group B and the other groups (P lt; 0.05), except mRNA expression of Desmin at 5 days between groups B and C, and mRNA expression of MyHC at 10 days between groups A and B (P gt; 0.05). Conclusion Low frequency (0.25 Hz) and suitable time (7 days) periodic mechanical stretch is beneficial to the differentiation of the multilayer myotubes cultured in the groove casts of Sylgard 184, but as the stretch time goes on the aging of myotubes will be accelerated.
Objective To investigate the role of preoperative serum thyroid stimulating hormone (TSH) in evaluating differentiated thyroid cancer (DTC). Methods A total of 551 patients with thyroid nodules met the study criteria, who got treatment in the First Affiliated Hospital of Jiamusi University between Aug. 2017 and Dec. 2017. And the patients were divided into DTC group (n=110) and benign group (n=441) according to the postoperative pathological results. The difference of serum TSH level between the 2 groups was compared and then explored the diagnostic significance of serum TSH level, thyroid imaging report and data system (TI-RADS), and serum TSH combined withTI-RADS. Results The serum TSH level was higher in the DTC group than that of the benign group (Z=5.198, P<0.05). The sensitivity of preoperative serum TSH level in the diagnosis of DTC was 80.9% (89/110), the specificity was 74.4% (328/441), and the area under receiver operating characteristic (ROC) curve was 0.660 ［95%CI was (0.602, 0.719), P<0.05］. The sensitivity of TI-RADS in the diagnosis of DTC was 82.7% (91/110), the specificity was 73.5% (324/441), and the area under the ROC curve was 0.772 ［95%CI was (0.711, 0.823), P<0.05］. The sensitivity of preoperative serum TSH combined with TI-RADS in the diagnosis of DTC was 91.8% (101/110), the specificity was 87.5% (386/441), and the area under the ROC curve was 0.831 ［95%CI was (0.786, 0.876), P<0.05］. Conclusions Preoperative serum TSH level may be a potential risk factor for the occurrence of DTC. Preoperative serum TSH level combined with TI-RADS classification can improve the accuracy of preoperative diagnosis of DTC, not only to reduce the misdiagnosis of thyroid cancer, but also can avoid excessive treatment.
ObjectiveIn order to improve the levels of clinical diagnosis and treatment of differentiated thyroid cancer, the research status and progress of blood markers of differentiated thyroid cancer in recent years were reviewed.MethodThe literatures about blood markers and liquid biopsy of differentiated thyroid cancer at home and abroad in recent years were searched and summarized.ResultsThyroglobulin and thyroglobulin antibody were the most commonly used for markers of differentiated thyroid cancer. The application value of blood markers such as microRNA and long non-coding RNA in the diagnosis, treatment and follow-up of differentiated thyroid cancer had also been found.ConclusionBecause of the advantages of high specificity, high sensitivity, and no-invasion, blood markers are useful indicators to help improve the diagnosis of thyroid cancer patients and monitor the disease progression and recurrence in the future.
OBJECTIVE: To explore the pluripotential and possible clinical application of adult stem cells. METHODS: The original articles on adult stem cells were extensively reviewed and the recent advances were summed up. RESULTS: Adult stem cells were located at different tissues of human beings and had the pluripotentiality of self-renewal and differentiation. Some adult stem cells, such as in marrow, nerve, muscle, fat, skin, liver, tissues, had the ability to differentiate into the unrelated cell type. CONCLUSION: The pluripotential, ubiquitous distribution and plasticity of adult stem cells offered a new way in regeneration medicine, such as cell therapy and tissue engineering.
OBJECTIVE: To investigate the biological characteristics of continuously subcultured human embryonic skeletal myoblasts, and choose the optimal seeding cells for muscle tissue engineering. METHODS: Human embryonic skeletal myoblasts were subcultured in vitro. The growth curve, rate of myotube formation(RMF) were used to evaluate the proliferative and differentiation ability of myoblasts, and to investigate the influence of fibroblasts contamination on myoblasts. RESULTS: The beginning 6 passages of myoblasts showed b proliferative and differentiation ability. From the 8th to 20th passage, the rate of fibroblasts contamination was increased, it mainly showed the growth characteristics of fibroblasts with increased proliferation and low differentiation. After subcultured to the 20th passage, the degeneration of myoblasts was obvious. CONCLUSION: The myoblasts within 6 passages should be used as the seeding cells of muscle tissue engineering because of b proliferative ability and high rate of myotube formation.
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.
ObjectiveTo systematically evaluate the reliability and stability of transoral endoscopic thyroidectomy vestibular approach (TOETVA) and conventional open thyroidectomy (COT) in the treatment of differentiated thyroid cancer.MethodsThe clinical studies of TOETVA and COT in the treatment of differentiated thyroid cancer were retrieved from major databases including PubMed, Embase, Cochrane Library, Wanfang, and CNKI by computer. The search date ended on March 1, 2020. Two investigators screened the literatures strictly and extracted the data following the pre-defined inclusion and exclusion criteria, and then used RevMan 5.3 software for meta-analysis.ResultsA total of 7 studies including 1 465 patients were included in this meta-analysis. The results showed: compared with the COT group, the operation time of the TOETVA group was longer [WMD=35.18, P=0.000 1], and the number of lymph node dissections in the central area was larger [WMD=1.42, P=0.000 5]. But the intraoperative blood loss [WMD=–5.32, P=0.39], the length of hospital stay after operation [WMD=0.05, P=0.94], the incidences of transient recurrent laryngeal nerve palsy [OR=0.81, P=0.43], transient hypocalcemia [OR=0.55, P=0.35], permanent hypocalcemia [OR=0.39, P=0.22], permanent recurrent laryngeal nerve palsy [OR=1.34, P=0.73], and hematoma [OR=1.29, P=0.69] were not statistically significant between the two groups.ConclusionsTOETVA has a higher stability. Although the COT has a shorter operation time, the former has a higher central lymph node dissection rate, and there is no scar on the neck after surgery and no significant difference in the incidence of postoperative complications.
Fracture is a common physical injury. Its healing process involves complex biological activities at tissue, cellular and molecular levels and is affected by mechanical and biological factors. Over recent years, numerical simulation methods have been widely used to explore the mechanisms of fracture healing, design fixators and develop novel treatment strategies, etc. This paper mainly recommend the numerical methods used for simulating fracture healing and their latest research progress, which helps people better understand the mechanism of fracture healing, and also provides direction and guidance for the numerical simulation research of fracture healing in the future. First, the fracture healing process and its relationship with mechanical stimulation and biological factors are described. Then, the numerical models used for simulating fracture healing (including mechano-regulatory model, biological regulatory model and mechano-biological regulatory model) and corresponding modeling techniques (mainly including agent-based techniques and fuzzy logic controlling method) were summarized in particular. Finally, the future research directions in numerical simulation of fracture healing were preliminarily prospected.