BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can be induced to differentiate into neuron-like cells in new environment after transplantation, and then to replace damaged cells for reconstructing neural circuit. OBJECTIVE: To establish the co-culture system between rat BMSCs and neural cells in vitro, and to study the influence of neural cells on the differentiation of BMSCs into neuron-like cells in the co-culture system. METHODS: The neural cells obtained from Wistar rat fetal brain tissue and BMSCs gained from rat thighbone were co-cultured in Transwell culture plate. The morphological changes of BMSCs were observed and the special markers of neural cells in BMSCs were examined by immunofluorescence on the fifth day of the co-culture. The results were compared with control group which where BMSCs were alone cultured. RESULTS AND CONCLUSION: BMSCs in the neural cells co-culture system extended, were radial, connected each other. Neuron specific enolase immunoreactions showed positive results, showing neuron-like cells. The positive ratio of neuron specific enolase-positive cells was (33.0±10.5)%. However, BMSCs in the control group did not express neuron morphological character. Immunofluorescence exhibited that cells were negative for neuron specific enolase. These indicate that microenvironment provided by neurons improves the differentiation of BMSCs into neuron-like cells.

Objective: To analyze the electrocardiographic (ECG) characteristics in patients with primary hyperparathyroidism (PHPT) and meanwhile, to explore the relationship between ECG abnormality and serum calcium level in relevant patients. Methods: Our study included 2 groups: Observation group,n=84 PHPT patients treated in our hospital from 2010-01 to 2014-04 and Control group,n=80 normal subjects from regular physical examination at the same period. The ECG manifestation was compared between 2 groups. Meanwhile, according to tertile levels of serum calcium, PHPT patients were further divided into 3 subgroups:①Ca2+ level (2.0-2.8) mmol/L,②Ca2+ level (2.8-3.1) mmol/L,③Ca2+ level >3.1 mmol/L andn=28 in each subgroup. The ECG abnormalities were compared among 3 subgroups and the relationship between ECG changes and serum calcium levels were studied. Results: The ECG abnormality rates in Observation group and Control group were as 83.3% vs 18.8%, P<0.001. Compared with Control group, Observation group presented more frequent ST-T changes, QRS duration widening, high left ventricular pressure and QT interval shortening,P<0.001 orP<0.05. In observation group, with the rising of serum calcium, the incidences of ST-T changes and high left ventricular pressure were increased accordingly, the ST-T changes among 3 subgroups were significant,P<0.05; while with the rising of serum calcium, the QT interval was shortened and QRS duration was widened accordingly, the differences were obvious among 3 subgroups, allP<0.05.Conclusion: PHPT patients have the higher incidence with abnormal ECG, while ECG abnormality has beenrelated to serum calcium level in relevant patients.

Objective To investigate the effect of manganese-enhanced MRI (MEMRI) at 7.0T for tracing nerve tracts in rat brain in vivo. Methods With brain stereotactic apparatus, 0.4 μl Mncl_2 with aqueous solution of 1 mol/L was injected into the right somatosensory cortex of 9 SD rats. MR scan was performed for tracing corticospinal tracts and other coherent nerve tracts pre-, and 24, 48, 72 h, 7 days post-injection with 7.0T micro-MRI system, respectively. Results Corticospinal tracts were showed in intact after Mn~(2+) administration from somatosensory cortex, thalamus, cerebral peduncle to pons at the time point of 24, 48, 72 h and 7 days, while the best tdisplaying was achieved at 24-48 h after Mn~(2+) administration. Simultaneously a small quantity of Mn~(2+) reached the opposite somatosensory cortex through the corpus callosum. Conclusion MEMRI for tracing rat nerve tracts can be showed clearly with 7.0T MRI. The location of manganese-enhanced corticospinal tracts in agreement with the rat brain atlas in stereotaxic is in agreement with that Paxinos' published. MEMRI can display the relationship between the two sides of hemisphere, and may play an important role in investigating the brain function and nerve plasticity after nerve injury in vivo.

Objective To study the cell vitality and ultra-structure of in vitro cultured fetus chondrocytes exposed to different doses of fluoride.Methods Primary chondrocytes were obtained from articular cartilage of the 24-27 weeks,aborted and dead fetuses.The third generation of primary cultured chondmcytes were exposed to concentrations of 0,10-2,5 × 10-3,10-3,10-4,10-5,10-6,10-7 and 10-8 mol/L fluoride for 24,48 and 72 h.Cell vitality was detected with Cell Counting Kit-8 (CCK-8) and ultra-structure of chondrocytes was observed by transmission electron microscope.Results The cell vitalities of chondrocytes exposed to doses of fluoride (10-2,5 ×10-3,10-3,10-4,10-5,10-6,10-7 and 10-8 moL/L) for 24,48 and 72 h were(15.04 ± 0.55)％,(62.53 ± 1.03)％,(100.34 ± 5.19)％,(111.40 ± 3.69)％,(121.47 + 6.09)％,(129.95 ± 4.96)％,(121.81 ± 4.97)％,(111.00 ± 1.63)％;(10.35 ± 0.64)％,(35.23 ± 2.41)％,(110.30 ± 2.07)％,(113.66 ± 6.98)％,(120.36 ± 6.23)％,(133.40 ± 5.80)％,(126.06 ± 5.40)％,(115.62 ± 7.33)％; (6.19 ± 0.16)％,(18.44 ± 0.21)％,(120.83 ± 4.93)％,(123.77 ± 4.82)％,(129.09 ± 5.21)％,(140.44 + 4.18)％,(131.99 ± 7.00)％,(124.10 ± 3.68)％,respectively.The cell vitalities of 10-2,5 × 10-3 mol/L fluoride groups were significantly lower than that of the control group (all P ＜ 0.05).The cell vitality of 10-2 mol/L group was significantly lower than that of the 5 × 10-3 mol/L group (P ＜ 0.05).Doses of fluoride (10-2,5 × 10-3 mol/L) could inhibit the cell vitality and promote the apoptosis of chondrocytes in vitro with increasing doses and prolonged time.The cell vitalities of 10-3,10-4,10-5,10-6,10-7,10-8 mol/L of fluoride groups were significantly higher than that of the control group (except the 24 h 10-3 mol/L,P ＜ 0.05).Between 10-4 and 10-3 mol/L groups(the vitalities of 48 h and 72 h were higher,but not significantly); 10-5 and 10-4 mol/L groups (the vitality of 72 h was higher,but not significantly); 10-6 and 10-5 mol/L groups,the cell vitalities were significantly higher than that of the control group(all P ＜ 0.05).Between 10-7 and 10-6 mol/L groups,10-8 and 10-7 mol/L groups (the vitality of 72 h was lower,but not significantly),the cell vitalities were significantly lower than that of the control group(all P ＜ 0.05).Doses of fluoride(10-3-10-8 mol/L) could promote the cell vitality of chondrocytes in vitro with prolonged time.The optimal concentration for the promotion was 10-6 mol/L.The cells of the control group were characterized as regular morphology,the abnormal surface microvillis,abundant cytoplasm and mitochondrial,abundant and slightly expanded rough endoplasmic reticulums and low electron-dense materials.The cells of 10-6 mol/L fluoride group had the following changes,increased and swell mitochondrial,hypertrophy and expanded rough endoplasmic reticulums.The cells of 5 × 10-3 mol/L fluoride group had the following changes,decreased microvillis,invaginated cell membrane,pyknosis and apoptotic body.Conclusion Doses of fluoride (10-3-10-8 mol/L) can promote the proliferation of human chondrocytes cultured in vitro.Doses of fluoride (10-2,5 × 10-3 mol/L) can promote the apoptosis of human chondrocytes cultured in vitro.

Salvianolic acid A (Sal A) is one of the most effective compounds isolated from the root of Salvia miltiorrhiza. Up to now, several studies regarding the pharmacokinetic profiles of Sal A have been reported, however there is no such study reported in monkeys, the species which is more similar to human. The aim of this study is to develop a LC-MS method for the determination of Sal A in monkey plasma and apply it to the pharmacokinetic studies of monkeys. After single intravenous administration of Sal A, the plasma concentration-time curves were observed and the main pharmacokinetic parameters were calculated. The plasma concentration at 2 min (C2 (min)) values were (28.343 ± 6.426), (45.679 ± 12.301) and (113.293 ± 24.360) mg x L(-1) for Rhesus monkeys treated with Sal A at 2.5, 5 and 10 mg x kg(-1). The area under the concentration-time curve (AUC(0-∞)) values were (3.316 ± 0.871), (5.754 ± 2.150) and (13.761 ± 2.825) μg x L(-1) x h, respectively. Furthermore, this method was improved and applied to the simultaneous determination of Sal A, Sal B and Sal C, which provided useful information for preclinical studies and clinical trials of Sal A, Sal B and Sal C.
Administration, Intravenous ; Animals ; Caffeic Acids ; pharmacokinetics ; Chromatography, Liquid ; Drugs, Chinese Herbal ; pharmacokinetics ; Lactates ; pharmacokinetics ; Macaca mulatta ; Mass Spectrometry ; Plant Roots ; chemistry ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo explore the procedures of dynamic infusion cavernosometry and cavernosography (DICC) and their application in the diagnosis and classification of venous erectile dysfunction (VED).

METHODSThis study included 103 ED patients, aged 20 to 43 years, highly suspected of VED, with disease courses of 4 months to 6 years. DICC was performed and analyses were made on the results, especially the parameters of flow-to-maintain (FTM) and pressure decay (PD) in the corpus cavernosum.

RESULTSBased on the parameters of FTM and PD, 21 of the patients were normal, 5 were suspected of VED, 39 had mild VED, 25 had moderate VED, and 13 had severe VED. Penile subcutaneous hematoma was found in 4 of the patients, all recovered after 3 to 5 days, with no other complications.

CONCLUSIONDICC is a reliable, safe and minimally invasive method for the diagnosis and classification of VED.

Diagnostic Techniques, Urological ; adverse effects ; Hematoma ; etiology ; Humans ; Impotence, Vasculogenic ; classification ; diagnosis ; Male ; Penile Diseases ; etiology ; Penis ; blood supply ; diagnostic imaging ; Radiography ; Veins

The paper is aimed to study the difference in yield and quality at different harvest time and determine the optimum harvest of planting Gentiana in Ludian traditional harvest period. The authors analyzed the variation in fresh weight, dry weight, dry discount rate, length, diameter, volume and the content of gentiopicroside, loganin acid, alcohol-soluble extract and total ash and made a comprehensive appraisal of yield, appearance quality and intrinsic quality by gray relational distance ideal Comprehensive Evaluation method. The results showed that there is a big difference in yield and quality both 2-year-old and 3-year-old Gentiana harvested in traditional harvest period and the comprehensive evaluation more better when harvested more later. It can be seen, Gentiana harvested the later had a better yield and quality in Ludian traditional harvest period. The harvest of Gentiana can be appropriate delayed depending on the particular circumstances of production.
Agriculture ; methods ; China ; Gentiana ; anatomy & histology ; growth & development ; metabolism ; Iridoid Glucosides ; metabolism ; Organ Size ; Quality Control ; Time Factors

Objective To synthesize N-(4-fluorophenyl)-N-(4-phenoxyphenyl) cyclopropane-1,1-dicarboxamide (NFNPDB) as small molecular c-Met kinase inhibitor analogue.Methods N-( 4-fluorophenyl )-N-( 4-phenoxyphenyl ) cyclopropane-1, 1-dicarboxamide was synthesized by nucleophilicsubstitution, amidation, etherification, reduction and condensation from diethyl malonate.Results The total yield of target compound was 3.79%, its structure was confirmed by 1 H-NMR.Conclusion The synthesis method of NFNPDB in our research can be easily operated with lost cost and short direction, which lays the foundation for designing the synthetic process of newly small molecular c-Met kinase inhibitor.

Salvianolic acid A (Sal A) is one of the most effective compounds isolated from the root of Salvia miltiorrhiza. Up to now, several studies regarding the pharmacokinetic profiles of Sal A have been reported, however there is no such study reported in monkeys, the species which is more similar to human. The aim of this study is to develop a LC-MS method for the determination of Sal A in monkey plasma and apply it to the pharmacokinetic studies of monkeys. After single intravenous administration of Sal A, the plasma concentration-time curves were observed and the main pharmacokinetic parameters were calculated. The plasma concentration at 2 min (C2 (min)) values were (28.343 ± 6.426), (45.679 ± 12.301) and (113.293 ± 24.360) mg x L(-1) for Rhesus monkeys treated with Sal A at 2.5, 5 and 10 mg x kg(-1). The area under the concentration-time curve (AUC(0-∞)) values were (3.316 ± 0.871), (5.754 ± 2.150) and (13.761 ± 2.825) μg x L(-1) x h, respectively. Furthermore, this method was improved and applied to the simultaneous determination of Sal A, Sal B and Sal C, which provided useful information for preclinical studies and clinical trials of Sal A, Sal B and Sal C.

Objective To study the role of manganese-enhanced MRI(MEMRI) in the depiction of cortical architecture of rat brain after systemic administration of Mn~(2+) through caudal vein and compare the effects of normal or opened blood-brain barrier on the manganese-enhanced MRI. Methods Fifteen SD rats were randomly divided into three groups according to ranked list of random. Blood-brain barrier was opened in short time by the injection of 30% mannitol via the right internal carotid artery, in group A, then 100 mmol/L MnCl_2 physiologic saline solution was delivered through vena caudalis, and MRI was performed 12 hours later. In group B, 100 mmol/L MnCl_2 physiologic saline solutions was administrated through vena caudalis, following normal saline injection into the right internal carotid artery, and MRI was performed 12 hours later. The group C served as normal control group. All images were acquired with a 7.0 T microMR scanner. Signal-to-noise ratios (SNR) in regions of interest were measured by Paravision 4.0 and the differences of three groups were compared by using one-way ANOVA. The differences of SNR on both sides of hemispheres were compared by using paired t test. Results MEMRI could show the gray matter and white matter of rat brain and the anatomy borders between somatosensory cortex and motor cortex clearly. Periventricular structures such as hippocampus, dentate gyms, habenula united, and olfactory bulb could also be showed clearly. Symmetrical enhancement on both sides of the cortex and banded structures was shown clearly in group B. The SNR increased and the differences were significant in right cerebral cortex, both sides of cerebellar cortex, hippocampus and pituitary, among three groups (right cerebral cortex 35.2±7.0,30.1±2.4,26.6±2.8,F =4.36,P=0.038;left cerebellar cortex 27.1±5.2,29.4±3.8,19.4±4.5, F=6.66, P=0.011;right cerebellar cortex 27.8±3.8,28.5±4.2,20.4±4.8, F=5.84, P=0.017; left hippocampus 34.5±4.9,38.1±5.3,24.5±3.6, F=11.38, P=0.002; right hippocampus 35.3±5.5, 37.6±4.7,25.6±3.0,F=9.93,P=0.003;pituitary 39.5±3.8,52.6±9.1,26.2±4.2,F=22.80, P=0.001) after systemic administration of Mn~(2+). Asymmetric enhancement on two sides of cortex was shown in group A. The mannitol-infused side was enhanced obviously but displayed blurring banded structures. However,the SNR differences of both sides of hemispheres in group A and B were not significant (P >0.05). Conclusions After systemic administration of MnCl_2 through vena caudalis, MEMRI could map the laminar architectures and the anatomy border of functional zone of somatosensory cortex specifically. High concentration of mannitol could open blood brain barrier(BBB) effectively and have distinct impacts on the architectures displayed in MEMRI. Opening or maintaining BBB in MEMRI had respective characteristics, and it should be selected according to practical needs.