BACKGROUND:Neural stem cells, as a hot topic in neuroscience research, have a wide application prospect in the treatment of neurological damage, but how to obtain a large number of terminally differentiated and purified nerve cells with homogeneous features is a difficult problem in this field. The use of intracellular fluorescence reporter system to track the process of neural stem cell differentiation and obtain a single kind of terminally differentiated and purified nerve cells provides a viable option. OBJECTIVE: To explore the value of GFAP promoter-driven fluorescence reporter system in tracing the neural differentiation of neural stem cells (NSCs). METHODS: Cerebral cortex of mouse embryos were primarily dissociated and sent for digesting and pipetting mechanically before suspension culture, followed by immunofluorescence staining of Nestin to identify their biological characteristics. Lentivirus carrying pLV/Final-neo-GFAP(promoter)-dTomato vector was employed to infect above-mentioned NSCs, and Geneticin (G418) was used to obtain purified NSCs at 14 days. Subsequently the purified cells were induced to differentiate into astrocyte-like cells; meanwhile red fluorescence changes in cells were observed by microscopy. The red fluorescent cells were then subjected to perform immunofluorescence staining at 13 days after induction. RESULTS AND CONCLUSION:The expression of Nestin in the isolated primary cells was strongly positive. Purified NSCs were obtained by lentivirus infection and subsequent G418 resistance selection at 14 days. After induced into astrocyte-like cells, the red fluorescence was observed in the cells under the microscope and furthermore, GFAP staining was also positive. Mouse NSCs carrying neo-GFAP(promoter)-dTomato were successfully obtained. The cells could express dTomato under the control of GFAP promoter, which provides a powerful tool for research on NSC differentiation mechanism, neural transplantation and tissue engineering product development.

BACKGROUND:As the pharmacological effect of eugenol constantly being discovered, its application in medical and food industry becomes wider. However, its toxicity studies have not established a complete database, especialy in the improvement of safety assessment of developmental toxicity and teratogenicity. OBJECTIVE:To establish a model of embryonic stem cel test to evaluate the embryotoxicity of eugenol. METHODS:Mouse fibroblasts (3T3) and mouse embryonic stem cels (E14TG2a) were culturedin vitro, and MTT test was performed to detect the cytotoxicity of 3T3 cels and E14TG2a cels with positive control 5-fluorouracil, negative control penicilin G and tested compound eugenol. The concentration of the tested compounds that inhibiting 50% viability of embryonic stem cels (IC50 E14TG2a) and 3T3 fibroblasts (IC50 3T3) was calculated. The hanging-suspension-adherent culture systems were used to induce embryonic stem cels into cardiomyocytes, and the concentration of tested compounds that caused 50% inhibition of differentiation of E14TG2a cels into cardiomyocytes (ID50 E14TG2a) was calculated. The embryotoxic potential of eugenol was classified by prediction model of the embryonic stem cel test. RESULTS AND CONCLUSION:The proliferations of E14TG2a and 3T3 cels were inhibited by eugenol, of which the IC50 3T3 and IC50 E14TG2a values were (3.613±0.192) and (1.799±0.131) mg/L. The differentiation of E14TG2a was also inhibited by eugenol, of which the ID50 E14TG2a was (3.501±0.158) mg/L. Eugenol was evaluated as a chemical compound with strong embryotoxicity by the model of embryonic stem cel test.

AIM: To develop a method to determine the content of emodin and chrysophanol in Fuyanling Effervescent Tablets(Radix et Rhizoma Rhei, Flos Lonicerae, Radix Sophorae Tonkinensis, etc.). METHODS: HPLC was used to determine the content of emodin and chrysophanol in Fuyanling Effervescent Tablets. The separation was performed on YWG ODS column with methanol (0.1%) phosphoric acid(85∶15) mixture as a mobile phase and the wavelength of UV detector was at 254nm. RESULTS: The resolution and the linearity of this method was good. in the range of emodin was 50.2～401.6ng( r =0.9999); chrysophanol was 49.9～ 399.2 ng( r =0.9994); and with the average recovery of emodin: 99.4%( RSD =2.2%); chrysophanol:100%( RSD =1.1%). CONCLUSION: The method is simple, rapid and satisfactory and suitable for quality control of Fuyanling Effervescent Tablets.

Objective To establish a model of embryonic stem cell test(EST)and utilize this model to evaluate the embryotoxici-ty of hydroquinone.Methods Mouse 3T3 fibroblasts and mouse embryonic stem(ES)cells(ES-E14TG2a)were cultured in vitro, and methyl thiazolyl tetrazolium(MTT)test was performed to detect the cytotoxicity of 3T3 cells and ES-E14TG2a cells induced by the positive control(5-fluorouracil),negative control(penicillin G)and tested compound(hydroquinone).The concentrations of the test compounds that inhibited 50% viability of ES-E14TG2a cells(IC50 ES)and 3T3 fibroblasts (IC50 3T3)were calculated.The hanging-suspension-adherent culture systems were used to induce embryonic stem cells into cardiomyocytes,and the concentrations of test compounds that caused 50% inhibition of differentiation of ES-E14TG2a cells into cardiomyocytes (ID50 ES)was calculated. The embryotoxic potential of hydroquinone was classified by prediction model of the embryonic stem cell test.Results The prolif-eration of ES-E14TG2a and 3T3 cells were inhibited by hydroquinone,of which the IC50 3T3 and IC50 ES values were (5.97±0.48) and (2.57±0.10)μg/mL respectively.The differentiation of ES-E14TG2a cells were also inhibited by hydroquinone,of which the ID50 ES was (3.77±0.31)μg/mL.Hydroquinone was evaluated as a strong embryotoxicity chemical by prediction model of EST. Conclusion Hydroquinone exhibits a strong embryotoxicity.

Objective:To evaluate the efficacy of Epstein-Barr nuclear antigen 1/immunoglobulin A (EBNA1/IgA), BamH1 Z transactivator/IgA (Zta/IgA), capsid antigen/IgA (VCA/IgA), and Epstein-Barr virus deoxyribonucleic acid (EBV-DNA) in detecting different stages of na-sopharyngeal carcinoma (NPC). The relationship between the EBV markers and stages of NPC was also analyzed. Methods:Blood sam-ples of 152 untreated patients with NPC and 675 healthy subjects were collected.ELISA was used to detect the serum levels of EBNA1/IgA, Zta/IgA, and VCA/IgA. Fluorescence quantitative PCR (FQ-PCR) was used to detect the plasma levels of EBV-DNA. ROC and correla-tion analyses were employed to assess the detection assays for NPC diagnosis. The positive rates of EBV markers in NPC patients in dif-ferent stages were analyzed statistically. Results: The positive rates of EBNA1/IgA, Zta/IgA, VCA/IgA, and EBV-DNA in NPC patients were higher than those in the healthy individuals. The expression of EBNA1/IgA was relatively high in early NPC. The sensitivity of EB-NA1/IgA was 77.8%. In advanced NPC, the level of EBV-DNA was high, and the sensitivity of EBV-DNA was 88.8%. The specificity of EBV-DNA and EBNA1/IgA could reach more than 96%. The combination of EBV-DNA and EBNA1/IgA showed the best diagnostic value, with a sensitivity of 92.1%(early stage 82.5%, advanced stage 98.9%) and a specificity of 96.9%. The positive rates of EBV-DNA were positively associated with the NPC clinic stage and N stage. The positives rates of Zta/IgA were positively associated with the NPC N stage. Conclusion:The best single index for NPC screening in an asymptomatic population is EBNA1/IgA. EBV–DNA is an ideal index for auxiliary diagnostics of advanced NPC. The combination of EBV-DNA and EBNA1/IgA shows the best diagnostic value. EBV-DNA is an important index in the stage and illness monitoring of NPC. Zta/IgA can indirectly reflect the character of lymph node metastasis, and it may be useful in assessment of NPC surveillance.

Objective To analyze the spatial distribution patterns of rodent community and the risk factors of disease transmission in natural foci of Xiji County,Ningxia,in order to provide a scientific basis for disease control and prevention.Methods Rodents were captured at different habitats in different latitudes which were selected with a stratified sampling method in 2012-2013.Capture rate of different rodent species and spatial distribution patterns of the animal community were analyzed.Antigen and antibody of hantavirus were detected in lung tissue and blood of the rodents with immunofluorescence and enzyme-linked immunosorbent assay (ELISA),respectively.Serum F1 antibody of Yersinia pestis (Y.pestis) was detected with indirect hemagglutination test and Y.pestis from liver and spleen tissue was cultured in vitro.Results Fourteen species of rodents were captured,belonging to 2 orders,7 families and 9 genera.Among them,ground squirrels and wood mouse were dominant species,accounted for 49.56％ (112/226) and 34.51％ (78/226) of the community,respectively.Infections of hantavirus and Y.pestis were not found in the rodent's community.Conclusion With the improvement of ecological environment in Xiji County,the spatial distribution patterns of rodent community is changing; the risk of zoonotic plague is reduced,but the risk of hemorrhagic fever with renal syndrome is being.

Physiological monitoring devices in modern clinical area are basically used electrodes or sensors directly touching the surface of human subject body, which will increase physiological and psychological load of the subjects. In order to realize non-contact monitoring of respiration and heartbeat, firstly, the micro bioradar was used to detect human body motion signal. Then, the respiration signal and heartbeat signal was extracted from the body-motion signal by using signal and conditioning circuits, digital filter and signal processing. Finally, the results of heart rate and breathing rate was wirelessly transmitted. The experimental results showed that the device for non-contact monitoring of respiration and heartbeat waveforms has advantages of small volume, low power consumption, which can realize the monitoring of physiological parameters in real time.
Heart Rate ; Humans ; Monitoring, Physiologic ; instrumentation ; Respiration ; Signal Processing, Computer-Assisted

Objective To explore the isolation,amplification methods and adipogenic differentiation under specific culture medium of human keloid-derived precursor cell (KPC) in vitro,in order to study their possibility of being new seed cells of tissue engineering fat.Methods KPCs were isolated from human keloid tissue of 4 different patients in our hospital and were cultured in the modified L-DMEM culture medium.Their cloning efficiency and growth curve were tested.The subcultured cells were tested of the mesenchymal stem cell (MSC)-related gene expression by flow cytometry.In addition,they were cultured in H-DMEM medium (containing 1 μmol/L dexamethasone,0.5 mmol/L 3-isobutyl-1-methyl-xanthine 10 mg/L of bovine insulin,100 mmol/L indomethacin,and 10 ％ FBS)and were later observed in oil red O staining under phase contrast microscope to determine whether lipid droplets generation was formed,using skin-derived precursors (SKP) as control.Results More than 95 ％ KPC expressed many antigens of MSC,such as CD29,CD44,CD90 and CD105 while few of them expressed CD34,CD45(1.0 ％-2.5 ％).And the cells increased in size gradually after inducted the same time,changing from spindle into round or polygonal in shape.The lipid droplets were seen in 72 hours and expressed a positive rate of 78.6 ％ in Day 19 in oil red O staining while the same rate was 54.6 ％ in SKP.Conclusions Human keloid-derived precursor cells can express a variety of MSC-related surface markers without expressing hematopoietic stem cell (HSC) related markers.Furthermore,they can be differentiated into fat cells under certain conditions,which may make them as a new source of seed cells for tissue engineering fat.

ABSTRACT:To improve the ability of correct classification and identification of rodents in different kinds of foci of zoonosis in Ningxia Hui Autonomous Region in China ,rodents samples from different habitats of Ningxia were collected .Sequences of COI gene were amplified and sequenced from 154 samples .Based on these sequences ,the pairwise genetic distance were calcu‐lated ,and a Neighbor Joining tree were constructed .According to the NJ tree ,20 clusters with high bootstrap support were found from 19 morphological species .The striped dwarf hamsters were divided into two clusters ,which suggested that there were two cryptic species with stripe on the back .The pika from Helan Mountain showed close relationship with Ochotona pal‐lasi ,and the genetic distance was as low as 3 .6% .Results show that DNA barcodes could be used to accurately identify speci‐mens of rodents and correct morphological identification errors .It could discover appearance indistinguishable implied species and could better study the classification and evolution of rodents .