Objective To investigate the biological effect of 50 Hz sinusoidal electromagnetic fields at 1 mT on the proliferation of the human osteosarcoma cell line MG-63.Methods Osteosarcrma MG-63 cells were divided into control and experimental groups.The control group was incubated without an electromagnetic field; the experimental group was incubated in a 50 Hz,1.0 mT sinusoidal electromagnetic field.On the 2nd,4th and 6th day,their proliferation was determined using a cell counting kit-8(CCK-8)assay.Variations in the cell cycle were detected with flow cytometry(FCM).Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)was used to measure cyclin B1 and cyclin D1 mRNA.Results Compared with the control group,proliferation of the experimental group cells was reduced significantly.The percentage of cells at G0-G1 phase increased,and the mRNA expression of cyclin B1 and cyclin D1 was significantly reduced.Conclusions A 50 Hz sinusoidal electromagnetic field at 1.0 mT can inhibit the proliferation of osteosarcoma cell line MG-63 significantly.

Objective:To investigate the risk factors of central lymph node metastasis (CLNM) in cN0 paillary thyroicl microcarcinoma (PTMC) and to establish a nomogram model for predicting the probability of cN0 PTMC CLNM.Methods:The clinicopathological data of 192 patients with cN0 PTMC admitted to the Department of General Surgery of the Second Hospital of Lanzhou University from Aug. 2016 to Aug. 2020 were retrospectively analyzed. There were 41 males and 151 females, 50 with CLNM and 142 without CLNM. The patients were divided into 2 groups according to the presence or absence of pathologically confirmed CLNM. Patient’s age, gender, tumor diameter, multiple, with Hashimoto’s disease, with nodular goiter, with or without near the posterior dorsal membrane, aspect ratio >1, with or without extratumoral infiltration, with or without lymphadenopathy, TSH levels, and TG levels were statistically analyzed. Pearson chi-square test was used to analyze the count data of hypothesis test, and the R language software package was used for Logistic multivariate analysis. The entry conditions were screened by stepwise regression to establish a nomogram prediction model, and the Bootstrap method was used for model verification. P<0.05 was considered statistically significant. Results:Multivariate logistic analysis showed that extratumoral invasion ( P=0.032) , presence of lymphadenopathy ( P=0.010) , and TG>68 μg/L ( P=0.007) were risk factors for central lymph node metastasis. The optimal model was established by stepwise regression. The factors included tumor diameter ≥0.5 cm, nodular goiter, extratumoral invasion, lymphadenopathy and TG>68 μg/L (AIC: 212.27) . The nomogram model was established according to the above risk factors. The consistency index (c-index) was 0.711. The results of calibration graph drawing and internal and external validation demonstrated its good consistency and applicability. Conclusion:Extratumoral invasion, lymphadenopathy, and TG>68 μg/L are risk factors for cN0 PTMC CLNM, and the nomogram established in the study can effectively predict the CLNM rate in patients with cN0PTMC and contribute to clinicians’ diagnosis and treatment decisions.

BACKGROUND: The source of bone marrow mesenchymal stem cells (BMSCs) is limited, and the cellular morphology,proliferation and multi-directional differentiation capacities can vary during serial passages in BMSCs in vitro.OBJECTIVE: To study the effects of fibroblast growth factor (FGF) on cellular morphology, proliferation and differentiation of serially passaged BMSCs.METHODS: (1) BMSCs were isolated from Sprague-Dawley rats and cultured. These cells were passaged six times in vitro, and the cellular morphology was observed and photographed. (2) BMSCs at passage 6 were seeded into 96-well plates and randomly divided into control group and FGF treatment group. The proliferation of cells in both groups was detected with cell counting kit-8 kit at days 1, 2, 3, 4, 5, 6, 7 after culture. (3) BMSCs at passage 6 were seeded into 6-well plates and randomly divided into control group and FGF treatment group. After 7 days treatment with growth medium or growth medium containing FGF, the cellular morphology was observed and photographed. And then the cells of both groups were treated with osteogenic induction medium, adipogenic induction medium and chondrogenic induction medium for the next 7 days. The osteogenic, adipogenic and chondrogenic related genes (RUNX2, ALP, OCN;PPARγ2, AP2, ADIPOQ; SOX9, collagen II, aggrecan) were detected with real-time PCR. The protein expressions of RUNX2, PPARγ2, SOX9 were detected with western blot assay. (4) BMSCs at passage 6 were seeded into 6-well plates and randomly divided into control group and FGF treatment group. After 7 days treatment with growth medium or growth medium containing FGF, the cells were cultured with osteogenic induction medium, adipogenic induction medium and chondrogenic induction medium for the next 14 days. Then, alizarin red S staining, oil red O staining and alcian blue staining were performed.RESULTS AND CONCLUSION: (1) After in vitro passage for six times, the cellular morphology changed obviously, and FGF treatment recovered the characteristics of primary cells. (2) Compared with the control group, the cell proliferation in the FGF treatment group was significantly increased (P < 0.05). (3) Compared with the control group, the expression of osteogenic, adipogenic and chondrogenic related genes (RUNX2, ALP, OCN; PPARγ2, AP2, ADIPOQ; SOX9, collagen II, aggrecan) was increased significantly in the FGF treatment group (P < 0.05). The protein expressions of RUNX2,PPARγ2, SOX9 were also higher in the FGF treatment group than the control group (P < 0.05). (4) Compared with the control group, the number of extracellular calcium nodules, the number of intracellular lipid droplets, and the expression of acid acidic mucopolysaccharide were significantly increased after FGF pretreatment. To conclude, FGF pretreatment can preserve the stemness of BMSCs serially passaged in vitro.

Objective To explore in vitro the best time window for using sinusoidal electromagnetic fields to promote the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).Methods BMSCs were isolated and cultured from 4-week-old Sprague-Dawley rats (male and female,80-120g).The BMSCs (from passage 3) were exposed 0,1,4 or 8h/d for 7d,14,or 28d,respectively,to 15Hz sinusoidal electromagnetic fields with a maximum amplitude of lmT.Those exposed 0h/d served as the control.The relative expressions of runt related gene-2 (RUNX2),bone sialoprotein (BSP) and osteopontin (OPN) were determined using real-time,quantitative reverse transcription-polymerase chain reactions (RT-PCRs).The level of RUNX2 protein was determined by Western blotting after 14d.Alizarin red staining was used to compare calcium distribution in each group.Results Obvious promotion of differentiation to osteoblasts was observed after 7 days of exposure to the15 Hz sinusoidal electromagnetic fields,most obviously manifested by an outstanding increase of the early osteogenic index RUNX2 in those exposed 4h/d.After 14 days of intervention,the 1h/d exposure showed to be most effective,especially in inducing the changes of the late osteogenic index OPN.The trends of changes in RUNX2 protein were similar in all groups.After stimulating 1h/d for 14 and 28days,calcium deposition increased to the greatest extent.Conclusions Exposure to sinusoidal electromagnetic fields induces osteogenic differentiation to osteoblasts in rat BMSCs in vitro.There is an apparent window effect.The best results are observed with more days of exposure and shorter exposure time (1h) every day.

BACKGROUND:Previous animal studies have revealed that osteogenic growth peptide (OGP) applied locally or systemically could promote fracture healing. But the disadvantages of short in vivo half-life and low oral bioavailability limit its clinical application.OBJECTIVE: To study the encapsulation and delivery of synthetic OGP (sOGP) from biodegradable polymeric microspheres in vitro so as to choose better carrier for the future study.DESIGN: Grouping observation and comparative trail.SETTING: Laboratory of School of Life Science and Technology, Xi'an Jiaotong University.MATERIALS: sOGP was synthesized by Xi'an Langene Bio-science Co., Ltd. with Fmoc system. The purity of sOGP after purification was over 98 % identified by reverse phase high performance liquid chromatography, and the molecular weight of sOGP was 1 523 650, which was consistent with the theoretical value (Mr 1 523 750); the result of whole sequence analysis of sOGP was consistent with the theoretical sequence of OGP. Poly (lactic-co-glycolic acid) (PLGA,50:50, Mr 30 000; 75:25 Mr 80 000) was obtained from Shandong Medical Instrumental Institute (Ji'nan, China)METHODS: PLGA with a 50:50 or 75:25 lactide to glycolide ratio was used for microsphere preparation using a modified double emulsion solvent extraction Water-in-oil-in-water (w/o/w) technique. The surface structure and appearance of microsphere was observed under scanning electron microscope; particle size distribution of microsphere was counted by laser diffraction particle sizer; efficiency of encapsulation, release time and the structural integrity of sOGP released from PLGA were assessed using high performance liquid chromatography (HPLC).RESULTS: ①Spherical microspheres of sOG-PLGA were formulated successfully. The average particle diameter of the PLGA 50:50 microsphere was (19.6±4.5) μm, efficiency of encapsulation (83.9±4.2)% with (83.9±4.2) % drug-loading efficiency, while the PLGA 75:25 microspheres showed an average size of (35.8±3.6) μm, efficiency of encapsulation (65.6±6.8)% with (65.6±6.8)% drug-loading efficiency. ②HPLC results indicated that sOGP were not chemically altered,physically aggregated but presented a intact structure as the original sOGP. An initial burst release was observed for both PLGA microspheres, especially from PLGA 75:25. sOGP was released from PLGA 50:50 microsphere for 56 days,and from PLGA 75:25 microspheres for over 70 days. The cumulative release of sOGP from PLGA 50:50 for 35 days was significantly lower than from PLGA 75:25 (P ＜ 0.05).CONCLUSION: The controlled release of sOGP encapsulated within PLGA 50:50 is better than the delivery from PLGA 75:25. Moreover, the release time could meet the requirements for fracture or bone defect site.

Objective To explore the effects of electromagnetic fields (EMFs) on osteogenesis during co-culture of bone mesenchymal stem cells (BMSCs) with osteoblasts in rats.Methods BMSCs and osteoblasts were isolated from Sprague-Dawley rats and cultured.Sub-cultured osteoblasts and BMSCs were seeded in transwell cell-culture-chamber polyester inserts to establish the co-culture system.The co-cultures were then randomly divided into a normal co-culture group and a group exposed to an EMF.Single-cultured BMSCs and osteoblasts were set as a single culture group.The EMF group was exposed to an EMF for 4 hours per day.On the 14th day,cell culture plates or inserts were randomly selected for total RNA extraction and measurement of the mRNA expression levels of Runt-related transcription factor 2,transcription factor 7,alkaline phosphatase,collagen type Ⅰ,bone morphogenetic protein 2 (BMP-2) and bone gamma-carboxyglutamate protein (Osteocalcin gene,OC gene) using real-time PCR assays.Cell culture dishes or inserts were also randomly chosen for Alizarin red staining to detect mineralized nodules.Results The level of osteogenic gene expression in single-cultured BMSCs and osteoblasts was low,while it was much higher in the co-culture group.The level of gene expression in the EMF-exposed and co-cultured group was even higher.Alizarin red staining also showed that calcium mineralized modules had increased in the stimulated,co-cultured system compared with the unstimulated,co-cultured cells.Conclusion EMF exposure can promote osteogenic differentiation of BMSCs and osteoblasts when they are co-cultured.BMP2-mediated cellular interaction might play an important role in osteogenic differentiation induced by EMF exposure.

Hepatorenal syndrome (HRS) is a common complication of severe liver disease, with severe conditions and poor prognosis, and causes a great burden to both patients’ family and society. HRS has a complex pathogenesis, and Western medicine treatment has a limited therapeutic effect; therefore, integrated traditional Chinese and Western medicine therapy is a feasible treatment method with a good clinical effect. This article reviews the advances in the diagnosis and treatment of HRS in both modern and traditional medicine, so as to overcome this challenge as early as possible.

OBJECTIVE: To investigate the diagnostic value of transcranial Doppler ultrasound in early stage cerebral arteriosclerosis. METHODS: We selected 50 patients with early cerebral arteriosclerosis as the disease group. At the same time we selected another 50 patients as a control group with no significant symptoms in the nervous system. By 2 MHz pulse Doppler probe through double-temporal windows and pillow windows Basilar artery (BA), the bilateral middle cerebral artery (MCA) was detected. In the TCD spectrum, we selected the spectrum of a single-family cardiac cycle, identified the starting point (ts), pulse-incisure point (ti), end point (te), and the peak velocity (tp), measured the time of the spectrum starting point to the peak velocity (Tp) and calculated the time required for the peak velocity in the share of ventricular systolic (Tp/Ti), the time required for the peak velocity in the share of the whole cardiac cycle (Tp/T). Tp, Tp/Ti and Tp/T were respectively named as time to peak velocity (TPV), peak-time index-1 (PTI-1) and peak-time index-2 (PTI-2). All data were analyzed by SPSS13.0. RESULTS: There was no significant difference in blood vessel velocity, PI, RI and S/D of BA and RMCA (P>0.05) between the control group and the disease group. Compared with the control group, TPV of the BA, LMCA and RMCA significantly extended, PTI-1 and PTI-2 of BA, LMCA and RMCA increased significantly in the disease group (P<0.01). In the disease group, there was no significant correlation between peak time index and PI, S/D (P>0.05). CONCLUSION TPV, PTI-1 and PTI-2 are sensitive indicators of early stage cerebral arteriosclerosis.
Adult ; Basilar Artery ; diagnostic imaging ; Blood Flow Velocity ; physiology ; Case-Control Studies ; Female ; Humans ; Intracranial Arteriosclerosis ; diagnostic imaging ; Male ; Middle Aged ; Middle Cerebral Artery ; diagnostic imaging ; Ultrasonography, Doppler, Transcranial

OBJECTIVE: To analyze cerebral hemodynamic changes by means of transcranial Doppler (TCD) and generally to explore the clinical application of this method in hyperlipemia patients. METHODS: Cerebral hemodynamics were detected by TCD in 63 patients with hyperlipidemia and compared with the hemodynamics of 64 health people. RESULTS: No statistically significant changes were found in the cerebral artery blood flow velocity and pulsatility index between the hyperlipidemic and control groups (P>0.05). Spectral shape, however, was abnormal in 52 patients in the hyperlipemia group (82.54%), which was statistically different (P<0.005) from controls. These abnormalities were classed as follows: 22 patients had abnormal spectra of the vertebrobasilar system, 2 patients had abnormal spectrum of the internal carotid arterial system, and 28 patients had abnormal spectra of both systems. The incidence of the abnormal spectra in vertebrobasilar system was significantly higher than the internal carotidartery (P<0.005). CONCLUSION TCD examination can reveal abnormal spectral shape in the cerebralartery and vertebrobasilar arterial systems in hyperlipidemia patients, and thus has some clinical value in determining changes in the brain of patients with high cholesterol levels and atherosclerosis.
Adult ; Cerebrovascular Circulation ; physiology ; Female ; Hemodynamics ; physiology ; Humans ; Hyperlipidemias ; diagnostic imaging ; physiopathology ; Male ; Middle Aged ; Ultrasonography, Doppler, Transcranial

Objective To explore the changes of plasma level of adrenomedullin(ADM)in congenital heart disease patients with heart failure,analyze the relationship between ADM and the severity of heart failure, assess the value of plasma ADM in the diagnosis of CHF and evaluate the cardiac function in pediatric patients with congenital heart disease. Methods 46 patients with CHD were enrolled according to the modified Ross Score. They were divided into three groups:mild CHF group(n = 16)and moderate CHF group(n = 18)and severe CHF group(n = 12). 25 matched normal children were enrolled as controls. Plasma ADM was measured with radioimmunoassay(RIA)and the contents of NT-proBNP were determined among all the patients. LVEF and E/A in all of them were also detected. The relationship between plasma ADM level and modified Ross Score and echocardiographic cardiac functional indexes was analyzed. Results Plasma ADM was positively correlated with modified Ross Score(r = 0.65,P < 0.01). Plasma ADM concentration in the severe CHF group was significantly higher than that of mild and moderate CHF group(P < 0.01),and plasma ADM concentration in the mild CHF group was higher than that in the control group(P<0.05).Plasma ADM was negatively correlated with LVEF and E/A(r =-1.05,P < 0.01). Conclusion Plasma ADM level could be used to assess cardiac function and diag-nose CHF with CHD.