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OBJECTIVE：To investigate th e effects of juglone on brain tissu e of rats and its relationship with the biomarkers related to brain tissue injury. METHODS ：Totally 40 rats were divided into blank group ，juglone high-dose ，medium-dose and low-dose groups （34.832，17.416，8.708 mg/kg），with 10 rats in each group. They were given medicine intragastrically once a day ， for consecutive 4 weeks.After last administration ，the general behavior ，brain index and brain tissue morphology were investigated. UPLC-MRM-MS method was used to determine the serum contents of L-dopa（L-Dopa），L-tyrosine（L-Tyr）and L-tryptophan（L-Trp） in rats. The chromatographic condition included Waters Acquity UPLC BEH C 18 column，mobile phase consisted of ammonium acetate aqueous solution-acetonitrile ，gradient elution ，at the flow rate of 0.3 mL/min，sample size of 5 μL，column temperature of 30 ℃；MS condition include electrospray ion source ，in positive ion mode ，capillary voltage of 3 500 V，desolvent gas flow of 650 L/h，desolvent temperature of 350 ℃，ion source temperature of 110 ℃. RESULTS ：Compared with blank group ，the rats in each dose group showed the behavior of tiredness and weakness of limbs. The brain tissue morphology showed pathological changes ， which contained blood vessel congestion in the cerebral and cerebellar cortex ，partial cell nucleus pyknosis in the pyramidal cell layer，deep staining of nuclei ，irregular shape and unclear boundary and other pathological changes ；the brain index of juglone high-dose group increased significantly （P＜0.05）. The established UPLC-MRM-MS method showed good specificity and the range of L-Dopa，L-Tyr and L-Trp were 31.25-32 000，31.25-32 000，15.625-16 000 ng/mL（r＝0.999 1-0.999 9），respectively. The limits of detection were 6.250，5.625，3.125 ng/mL，respectively. The limits of quantitation were 15.625，18.75，10.00 ng/mL，respectively. RSDs of precision ，accuracy and stability （24 h）tests were all Matrix effects were 95.1％-100.1％ （RSD are not more than 3.25％，n＝3）. Compared with blank group，the contents of L-Dopa were increased significantly injuglone medium-dose and high-dose groups （P＜0.01）. The contents of L-Tyr were increased significantly in juglone lowdose，medium-dose and high-dose gro ups，while the contents of L-Trp were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：Juglone has an effect on the general behavior，brain index and brain tissue morphology of rats. It may affect the brain function of rats by increasing the contents of L-Dopa and L-Tyr in serum and decreasing the contents of L-Trp.
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Objective@#To investigate the effect of siRNA silencing α1 antitrypsin (α1-AT) gene on the biological behavior of rheumatoid arthritis fibroblast-like synoviocytes (RA-SFs).@*Methods@#Primary culture of knee synovial tissue from 5 patients with rheumatoid arthritis (RA) was performed. The artificially synthesized silencing α1-AT siRNA specifically inhibits the expression of α1-AT in RA-SFs. After 24 and 36 hours of transient transfection, the inhibition efficiency was detected by Quantitative reverse transcription polymerase chain reaction (RT-qPCR), and the expression of related genes after α1-AT gene silencing was detected.Furthermore, ethyl thiazolyl tetrazolium (MTT) assay, Trans-well chamber, cell scratch and enzyme-linked immunosorbent assay (ELISA) were used to detect the effects of interfering α1-AT expression on cell proliferation, invasion and migration, and secretion of interleukin (IL)-17, Tumor necrosis factor (TNF)-α, IL-1α, IL-1β and other related inflammatory factors. At the same time, when the pathway inhibitor (ERK inhibitor, signal transducer and activator of transcription 3 (STAT3) inhibitor, NF-κB inhibitor) stimulated cells, the effect on α1-AT was changed. One-way analysis of variance was used for comparison between the two groups; further pairwise comparison using LSD-t test; The count data was checked by χ2 test.@*Results@#Compared with the negative control group, the siRNA α1-AT group was transfected for 24 hours, the α1-AT mRNA level was significantly inhibited (P<0.05), and the proliferation rate of RA-SFs was not affected (P>0.05), and the invasion and migration ability of cells decreased significantly (P<0.05). The secretion of IL-17 and IL-1β was slightly decreased (P>0.05), while TNF-α and IL-1α were not affected (P>0.05). In addition, the expression of α1-AT downstream genes Matrix metallop roteinases (MMP)-2 169, Hu-Bax1, MMP-9, Hu Bax and Hu Bcl-xl were significantly decreased (P<0.05). Moreover, the signaling pathway inhibitors ERK inhibitor (PD98059) and NF-κB inhibitor (PDTC) had significant effects on α1-AT (P<0.05).@*Conclusion@#The α1-AT gene silencing has potential effect on the biological behavior of RA SFs. Further study of this mechanism is helpful to provide evidence for the treatment of RA.
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OBJECTIVE： To identify chemical components of Forsythia suspense. METHODS： UPLC-Q-TOF-MS technology was used for the chemical components analysis of F. suspense. The determination was performed on ACQUITY UPLC BEH C18 column with mobile phase consisted of 0.1％ formic acid aqueous solution （A）- 0.1％ formic acid acetonitrile solution （B） with gradient elution， at the flow rate of 0.3 mL/min； the column temperature was set at 30 ℃； the sample size was 5 μL. Positive and negative ions were detected by electrospray ionization. The temperature of the ion source was 550 ℃； the atomizing gas was N2； the atomizing gas and the auxiliary pressure were 379.2 kPa； the air curtain pressure was 241.3 kPa； the decluster voltage was 80 V/－80 V； the collision energy was 35 eV/－35 eV； the mass scanning range was 80-1 500 Da. Peakview 2.0 software was used to screen the target components by the first-order mass spectrometry， and calculate the high-resolution and accurate molecular weight of each component， compare with the reference spectrum and related literature， or calculate the elemental composition of fragment ions in the second-order mass spectrometry， analyze their decomposition pathways， then infer the structure of compounds. RESULTS & CONCLUSIONS： 45 kinds of compounds were identified from F. suspense，which included 7 phenylethanol glycosides，5 lignans，5 terpenes， 12 flavonoids，7 organic acids，2 phenols，2 quinones，2 glycosides and 3 other components. There were 19 compounds identified for the first time in F. suspense. The study provides a reference for the in-depth study of the pharmacodynamic substance basis of F. suspense and the rapid qualitative and quantitative analysis of the components.
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OBJECTIVE： To analyze chemical components as coumarin in Saposhnikovia divaricata， and to provide reference for comprehensive analysis of pharmacodynamic material base in S. divaricata. METHODS： UPLC-Q-TOF-MS method was adopted. Chromatographic condition： the determination was performed on ACQUITY UPLC BEH C18 column with mobile phase consisted of 0.1％ formic acid water-0.1％ formic acid acetonitrile （gradient elution） at the flow rate of 0.3 mL/min. The column temperature was 35 ℃， and sample size was 3 μL. MS condition： ESI， in positive and negative ion mode， ESI+/ESI－ 5 500 V/－4 500 V， declustering potential of 80-－80 V， auxiliary heating gas pressure of 55.00 psi， atomizing gas pressure of －55 psi， curtain gas pressure of －35.00 psi， desolvent temperature of 550 ℃， collision activation scanning energy of 15 eV， collision voltage of 35 psi. Component analysis was performed by comparing with the related literature data and the standard chromatogram control combined with accurate relative molecular mass of compounds provided by UPLC-Q-TOF-MS. RESULTS： Totally 135 chemical components were analyzed in ESI+ mode， and the 105 chemical components were analyzed in ESI－ mode. 11 chemical components as coumarin were identified in ESI+ mode， such as isoimperatorin， umbelliferone， scopolamine， xanthotoxin， psoralen， Ostenol， fraxidin， isoimperatorin， 5-hydroxyl-8-methoxypsoralen， phellopterin， decursin. CONCLUSIONS： The method is accurate and rapid， and can be used for the analysis of chemical components as coumarin in S. divaricata.
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Objective To explore pre-attentive information processing in undergraduate students with major depressive disorder(MDD) indexed by auditory mismatch negativity (MMN) and provide scientific theory basis for the diagnosis of MDD.Methods In the deviant-standard reverse oddball paradigm with deviant stimulus presentation probability of 20％,duration auditory MMN was obtained in 17 MDD university students and 17 gender-/age-matched healthy undergraduate students.All data were analyzed by statistics means of repeated measurement of F test.Results The amplitudes of 150 ms MMN over the frontal-central area were smaller in MDD undergraduate students((-0.34± 1.24) μV) than those in healthy undergraduate students ((-1.07± 1.55) μV,F=24.318,P=0.000).The 50 ms MMN did not differ between MDD students ((-1.96± 1.14) μV) and healthy undergraduate students ((-1.23 ± 1.97) μV,F =0.089,P =0.767).Conclusion Pre-attentive auditory information processing is impaired in MDD undergraduate students.
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A silver needle-knife which has the dual function of silver needle and needle-knife is designed. The main components of this silver needle-knife are approximately 50% silver and approximately 50% nichrome. The silver needle-knife is composed of five parts, including needle-knife tail, spiral handle; steering handle, needle-knife body and needle-knife edge. It converges the advantages of needle-knife and silver needle, which can cut loose of diseased tissue and peel adhesion of lesions, but also be heated with moxa cone and thermal therapeutic instrument, and connect with electroacupuncture apparatus. It has the function of warming channel and removing coldness, dispelling wind and eliminating dampness, resolving spasm and relieving pain, dredging the channel and so on. Due to the spiral handle and the steering handle, the operation is easier, which reduces the blindness of cutting and increase the safety. It is mainly used for soft tissue injury, rheumatism and rheumatoid arthritis, as well as degenerative diseases of spine and joint, and it has obvious efficacy on some internal medical diseases.
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Humanos , Terapia por Acupuntura , Diseño de Equipo , Agujas , Estándares de ReferenciaRESUMEN
AIM:To explore an ideal method to induce the differen-tiation of human umbilical cord mesenchy-mal stem cells (hUCMSCs) into neuron-like cells and to provide some evidence for the transplantation of hUCMSCs for spi-nal cord injury .METHODS:The hUCMSCs were isolated from human umbilical cord digested with collagenase Ⅱ.The hUCMSCs was verified by flow cytometry analysis .The passage 5 cells were randomly divided into 4 groups.The differentiation of hUCMSCs was induced by bFGF in group A , bFGF and BDNF in group B, or BHA, bFGF and BDNF in group C, while the cells in group D served as a control group cultured with DMEM-F12 and 10%FBS.Two weeks later , the expression of nestin , neurofilament protein H ( NEFH) and glial fibrillary acidic protein ( GFAP) was detected by real-time PCR and immunocytochemistry .The morphological changes of cells were observed under an atomic force microscope . RESULTS:Mesenchymal stem cells were isolated and cultured from human umbilical cord by enzyme digestion .hUCMSCs expressed CD29, CD44 and CD105, but no CD34, CD45 or HLA-DR.After cultured with inducing medium for 2 weeks, the cells were successfully induced into neuron-like cells.The appearance of the cells had great change .The induced hUC-MSCs developed round cell bodies with multiple neurite-like extensions observed under an atomic force microscope .The re-sult of real-time PCR showed that nestin was positive in A , B and C groups , and NEFH was positive in A and B groups , but GFAP was negative in 4 groups.The difference of nestin and NEFH expression among the induced groups was signifi -cant (P<0.05).CONCLUSION:Mesenchymal stem cells were isolated and cultured from human umbilical cord by en-zyme digestion in vitro, and all the hUCMACs presented stable biological properties .Moreover, hUCMSCs were induced to differentiate into neuron-like cells in vitro via bFGF combined with BDNF .
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<p><b>OBJECTIVE</b>To investigate the effect of ganoderic acid A (GA-A) on the biological behaviors of human osteosarcoma cells in vitro.</p><p><b>METHODS</b>MG63 and HOS cells were treated with 0.1, 0.25, and 0.5 mmol/L GA-A, and the changes in cell proliferation, apoptosis and migration were evaluated using MTT assay, flow cytometry, and Transwell assay, respectively. The expressions of STAT3, p38, and NF-κB1 in the cells were analyzed by Western blotting.</p><p><b>RESULTS</b>GA-A effectively inhibited the proliferation of human osteosarcoma HOS and MG-63 cells in a dose-dependent manner, and induced obvious cell apoptosis in both cells. Treatment with 0.5 mmol/L GA-A also resulted in significant inhibition of the invasion of both cells. The results of Western blotting showed that GA-A down-regulated the expression level of phosphorylated STAT3 and increased the phosphorylation level of p38 and NF-κB1 expression in both cells.</p><p><b>CONCLUSION</b>GA-A can induce proliferation inhibition, apoptosis and suppression of invasion in human osteosarcoma HOS and MG-63 cells.</p>
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Humanos , Apoptosis , Neoplasias Óseas , Patología , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Ácidos Heptanoicos , Farmacología , Lanosterol , Farmacología , Subunidad p50 de NF-kappa B , Metabolismo , Osteosarcoma , Patología , Fosforilación , Factor de Transcripción STAT3 , Metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos , MetabolismoRESUMEN
Objective To evaluate the inverse distance weighted(IDW) in revealing the characteristics of spatial distribution of water fluoride in Heze City,Shandong Province.Methods A geographic information system (GIS) database of water fluoride was established in Heze City of Shandong Province using the data of endemic fluorosis surveys collected by Endemic Disease Prevention Institute in Shandong Province during 2005-2007.IDW spatial interpolation was applied to predict the distribution of fluoride in drinking water in 139 towns of Heze City.Sensitivity and specificity were calculated.Results Mean water fluoride levels in 10 counties of Heze City were all higher than 1.0 mg,/L,and the water fluoride in Cao County,Juye,Mudan District and Juancheng were higher than 2.0 mg/L.Of all 139 townships of Heze City,129 were higher fluoride townships where fluoride was ＞ 1.0 mg/L,10 were lower fluoride townships(≤ 1.0 mg/L).IDW spatial interpolation showed that the water fluoride of most areas in Heze City were ＞ 1.0-2.0 mg/L.The areas with water fluoride of ＞ 2.0-3.0 mg/L were mainly located in eastern Juancheng,northern Mudan District,north-central Chengwu,central and southern Juye,southeastern part of Caoxian and eastern part of Shan Town.Regions of water fluoride ＞ 3.0 mg/L were mainly distributed in Xieji and Wanfeng towns of Juye County,Jishan town of Juancheng County,Sunlaojia town of Caoxian and Dusi town of Mudan County.The internal verification results showed that the sensitivity,specificity and overall accuracy rate of IDW used for predicting water fluoride content was 100.00％ (129/129),10.00％ (1/10) and 93.53％ (130/139),respectively.While the external verification results showed that the sensitivity,specificity and overall accuracy rate of IDW for predicting water fluoride content was 100.00％(31/31),11.11％(1/9) and 80.00％(32/40),respectively.Conclusion With the application of IDW interpolation,it is feasible to infer the overall spatial distribution based on the monitoring data,and to reveal the spatial characteristics of water fluoride in Heze City,Shandong Province.
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Objective To investigate the epidemic status of endemic fluorosis in Shandong Province,Jining City,and to provide a basis for prevention and control of the disease.Methods Based on Shandong Provincial Project Technical Solutions for Endemic Fluorosis,Rencheng,Jinxiang,Yutai,Jiaxiang and Liangshan Counties in Jining were selected as monitoring sites.According to the illness situation of mild,moderate or serious districts,one village was selected as a major survey site from each county(district).There were a total of 15 such villages selected.Survey content included drinking water fluorine level; dental fluorosis of children,adults' clinical skeletal fluorosis and urinary fluorine levels; water and urinary fluoride content were determined by the method of fluoride ion selective electrode; dental fluorosis of children was diagnosed by Deans method and clinical diagnosis was based on the Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008).Results Sixty-one water samples from 15 villages of five counties (districts) were tested.Fluoride levels of 9 out of the 61 samples were exceeded the national standard (＞ 1.0 mg/L),and the rate was 14.75％; 1 sample ＞ 2.0 mg/L,and the maximum water fluoride was 2.25 mg/L.Seven hundred and seventeen people's real time urinary fluoride was detected in the 15 villages,including 420 children and 297 adults,and the geometric mean were 1.53 and 1.69 mg/L,respectively.Clinical examination of 755 children aged 8 to 12 showed that the detection rate of dental fluorosis was 26.89％ (203/755); defect rate was 9.12％(29/755) and dental fluorosis index weres 0.65.The detection rate of clinical skeletal fluorosis of 11 565 adults was 4.76％(550/11 565),including 303 moderate or serious cases.Conclusions The situation of excessive water fluorine in outside environment in Jining City has been controlled at a certain degree; groups urinary fluoride level is closed to the normal upper limit; the prevalence of dental fluorosis or skeletal fluorosis has been suppressed at a certain degree,therefore,the results of control should be further consolidated and expanded,in order to completely eliminate the fluoride hazard.
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Objective To investigate the situation of ALT-single-unqualified blood donors and the ALT test when they donated again.Methods There were 3 784 cases of ALT-single-unqualified blood donors from March 2009 to February 2010 enrolled in the study.Investigations were carried out to know the previous situation of blood donation.A 3-year tracking survey on those people was carried out,and the data was recorded and analyzed.Results The ALT-single-unqualified blood donors who participated in blood donation for the first time accounted for 58.14% (2 201/3 748).The 3-year follow-up showed that the returned blood donors accounted for 33.62% (1 272/3 784);1-year return accounted for 46.62%(593/1 272)which was the most;ALT-qualified donors accounted for 65.72%(836/1 272)of the retured donors.with the increase of the times of blood donation,the qualified rate of blood ALT increased.Conclusion More than half of the ALT-single-unqualified blood donors returned.There was a large proportion of returning donors participated in blood donation more than onece,and the qualification rate of ALT increased with the increase of do-nation times.In order to reduce the unqualified rate of ALT test,we should strengthen the propaganda and fixed blood donation team construction.
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Objective To analyze the assessment results of external quality control and network operation of urinary iodine in local laboratories of Shandong Province,to evaluate the ability of consistent analysis; and to provide a reliable laboratory quality assurance for epidemiological surveillance and control of iodine deficiency disorders (IDD) and reliable decision-making.Methods Z-scores of the assessment results of external quality control of urinary iodine in local laboratories of Shandong Province were analyzed from 2000-2013.Results The feedback rate of urinary iodine in local laboratories of ShandongProvince was 100.0％ from 2000-2013; the qualified rate was 100.0％(14/14),100.0％(14/14),92.9％(13/14),100.0％(14/14),92.9％(13/14),100.0％ (14/14),100.0％ (14/14),100.0％ (14/14),92.9％ (13/14),100.0％ (17/17),100.0％ (17/17),94.1％ (16/17),100.0％ (17/17),and 100.0％ (17/17),respectively.The total qualified rate of Z-scores between Zs in local laboratories was 100.0％ (214/214)and within Zs was 98.6％ (211/214).Conclusion The test abilities of urinary iodine in local laboratories of Shandong Province are steady; the overall results are satisfactory,but some laboratories need to improve.
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<p><b>OBJECTIVE</b>To evaluate the clinical effect of different surgical approaches for treating cervical ossification of the posterior longitudinal ligament (OPLL) with spinal cord signal change.</p><p><b>METHODS</b>Thirty-eight patients with OPLL with spinal cord signal change were treated from January 2005 to January 2011. Surgical removal via an anterior approach or partial decompression was performed in 10 cases (group A), posterior approach open-door laminoplasty with decompression, bone grafting and internal fixation was performed in 12 cases (group B), and opening the cervical spinal meninges to relieve the pressure was performed in 16 cases (group C) on the basis of the procedures in group B. All the patients were followed up and the pre- and postoperative JOA scores, improvement ratio and inter-body implant fusion were evaluated. Imaging examinations including X-rays, CT and MRI were also performed pre- and postoperatively, and the surgical complications were recorded.</p><p><b>RESULTS</b>At 12 months postoperatively, the mean improvement rates in groups A, B, and C were 52.39%, 55.15%, and 60.32%, respectively, with the mean JOA scores of 13.54∓0.56, 13.56∓1.26, and 14.70∓1.41, respectively. The JOA scores and improvement rates significantly increased after the surgeries. One patient in group A became paraplegic after the operation with cerebrospinal fluid leakage, and one patient in group B and one in group C reported numbness of the upper limb. Group C showed a shorter postoperative recovery time without severe complications.</p><p><b>CONCLUSION</b>Posterior open-door laminoplasty, decompression, bone grafting and internal fixation can be an effective approach for treatment of cervical OPLL with spinal cord signal change and requires shorter rehabilitation time after the operation.</p>
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Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Vértebras Cervicales , Patología , Descompresión Quirúrgica , Métodos , Osificación del Ligamento Longitudinal Posterior , Patología , Cirugía General , Compresión de la Médula Espinal , Cirugía General , Resultado del TratamientoRESUMEN
BACKGROUND: Percutaneous vertebroplasty for osteoporotic vertebral compression fractures has achieved very good results, but its long-term efficacy as well as impact on patients has been rarely reported so far.OBJECTIVE: To investigate the long-term effect of vertebroplasty with bone cement on osteoporotic vertebral compression fractures through a follow-up.METHODS: Thirty-four patients with osteoporotic vertebral compression fractures who had undergone percutaneous vertebroplasty were recruited. Visual analogue scale scoring was measured and compared as well as lesioned vertebral height and kyphosis angle shown on lateral X-ray examination prior to, 1 week and 6 years after percutaneous vertebroplasty.RESULTS AND CONCLUSION: The kyphosis angle was improved 1 week and 6 years after percutaneous vertebroplasty, and it changed insignificantly during the follow-up period. The vertebral height was also improved significantly after percutaneous vertebroplasty (P < 0.01); however, there was no obvious variation in the vertebral height at 1 week and 6 years after percutaneous vertebroplasty. The visual analogue scale exhibited an improvement after percutaneous vertebroplasty (P < 0.01); however, with time going by, the scoring on the visual analogue scale had an increased tend. All the parameters remained stable and had no large fluctuations. It is proved that the percutaneous vertebroplasty is effective and safe to treat osteoporotic vertebral compression fractures with an excellent long-term effect.
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Obje:ctive To establish an optimized method to isolate, culture and identify human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and induce their osteogenic and adipogenic differentiation. Methods The hUCMSCs were isolated from human umbilical cord by digestion with collagenase. After serial subcultivation in vitro, the stem cells were passaged. Morphologic appearance of hUCMSCs was observed under an optical microscope and atomic force microscope. The proliferation rate was measured by MTT assay. Cell cycle and surface antigens were measured by flow cytometry. The osteogenic and adipogenic differentiation was tested and evaluated by specific staining methods. Results The isolation of hUCMSCs by digestion with collagenase was efficient. After seeded for 24 hours, the adherent cells showed spindle shape and fibroblast cell-like shape and the size of hUCMSCs was homogeneous. The similar growth curves of passage 3 and 7 exhibited a great potential for proliferation. Flow cytometry analysis revealed that CD29, CD44 and CD105 were highly expressed on the surface of passages 3 cells, but the expression was negative for CD34, CD45 and HLA-DR. After culture in inducing medium, the cells were successfully induced into osteogenic and adipogenic lineages. These cells were highly positive for alkaline phosphate staining and also showed mineralization presented with von kossa staining after 4 weeks' culture induction of osteogenic differentiation. Furthermore, liquid vacuoles were detected by oil red O staining after 3 weeks' culture induction of adipogenic differentiation. Conclusion An in vitro method for isolation and purification of hUCMSCs from human umbilical cord has been established. The cultured cells were composed of only undifferentiated cells and their biological properties were stable. The hUCMSCs are expected to be a new type of stem cells of tissue engineering.
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BACKGROUND: Mesenchymal stem cells (MSCs) are commonly observed under the scanning electron microscope, transmission electron microscope and inverted microscope. However, above-mentioned observation methods have disadvantages on observing ultrastructure of cell membrane and cytoskeleton. OBJECTIVE: To establish a more effective and appropriate method to isolate, culture and identification of human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and to study the ultrastructure of osteogenic differentiation with the atomic force microscope. METHODS: The hUCMSCs were isolated from human umbilical cord by digested with collagenase. After serial subcultivation in vitro, the stem cells were passaged, and osteogenic differentiation was determined by alkaline phosphatase calcium-cobalt staining and alizarin red staining. hUCMSCs immunophenotype was measured by Flow cytometry.The membrane surface ultrastructure of osteogenic differentiation was observed by Atomic Force Microscope before and after induction. RESULTS AND CONCLUSION: The isolated hUCMSCs by digested with collagenase was efficient. After seeded 24 hours, the adherent cell showed spindle shape, polygonal shape and fibroblast-cell-like shape and the size of hUCMSCs was homogeneous. Flow cytometry analysis revealed that CD29, CD44, CD105 were highly expressed on the surface of passages 3 cells, but there was negative for CD34, CD45 and HLA-DR. These cells were high positive for alkaline phosphate staining and also showed significant calcium node using alizarin red staining after 4 weeks culture induction of osteogenic differentiation. Under atomic force microscopy, undifferentiated stem cells demonstrated insignificant microtubule protrution in a parallel distribution following analysis of cytoskeleton before and after differentiation.
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A mutation is predominant in weak D individuals,and DⅥⅢ mutation in partial D individuals.
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AIM Valproate (VPA) is widely used to treat epilepsy. Long-term treatment of women with VPA has been reported to be associated with a PCOS-like syndrome. The aim of the present study was to investigate the effects of clinically relevant concentrations of valproate on steroidogenesis and steroidogenesis acute regulatory protein (StAR),cholesterol side-chain cleavage enzyme (P450scc)and cytochrome P450 aromatase (P450arom) mRNA expression in human luteinized granulosa cells in vitro. METHODS Human luteinized granulosa cells were isolated from oocyte retrieval of in vitro fertilization procedure and were cultured with DMEM medium and treated with various concentrations of valproate (0, 100, 250 mg?L -1), the culture media was collected after 2 days for progesterone and estradiol measurements by standard radiommunoassay, StAR, P450scc and P450arom mRNA in granulose cells were detected by fluorescent real-time RT-PCR (TaqMan assay). RESULTS Valproate caused significant increase of progesterone (P