Satellite cells, as the major stem cells, are responsible for postnatal skeletal muscle growth, regeneration and maintenance. As a result, satellite cells have great potential as therapeutic agents. Activation of satellite cells in vivo is a key link in the muscle regeneration processes. And their replenishment is necessary to keep the capacity of skeletal muscle to regenerate after recurrence of muscle damage. Cellular and molecular regulation of activation and replenishment for satellite cells issues were reviewed. Hoping through the points of nitric oxide-hepatocyte growth factor (NO-HGF), myostatin and Notch signaling and the niche of satellite cells to overcome the recent obstruction in cell therapy in clinic myopathy, such as Duchenne muscle destrophy.

OBJECTIVEThe aim of this study is to establish a transgenic mouse model for cleft palate relevant gene thyroid transcription factor-2 (TTF-2), which can be used to study palatal shelf development when the expression pattern and regular activation of TTF-2 is altered.

METHODSThe C57BL/6J mouse TTF-2 gene was cloned through polymerase chain reaction (PCR) from the mouse genomic DNA. The TTF-2 gene was inserted into the expression vector pBROAD3-mcs to construct the recombinant expression vector pBROAD3-TTF-2. This expression vector was then microinjected into the male pronuclei of the fertilized mouse ovum. Thus, the TTF-2 transgenic mice model was established. The genotype of the transgenic mice was identified by PCR and Southern blot analysis. Immunohistochemistry identified the consistent expression of TTF-2 gene during its palatal shelf development.

RESULTSTTF-2 genes were microinjected into 982 fertilized ova. A total of 580 two-cell-stage embryos cultured and transplanted into the oviducts of 48 pseudopregnant female mice. Overall, 68 embryos were obtained for analysis. The genotype of the mice was determined through PCR and Southern blot analysis using genomic DNA extracted from tail biopsies of the transgenic fetus. A total of 13 TTF-2 transgenic mice were detected. The expression of TTF-2 gene during the palatal shelf development of the transgenic mice was consistently detected by immunohistochemistry.

CONCLUSIONThe recombinant expression vector pBROAD3-TTF-2 was integrated into mouse genome through microinjection. The transgenic mouse in the palatal shelf that consistently expressed TTF-2 was successfully established and displayed a cleft palate phenotype.

Animals ; Cleft Palate ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Thyroid Gland

Objective To identify the Ligustrum sinense Lour.and Jasminum elongatum(Bergium) Wild.from the macroscopic appearance and microscopic features.Methods Fresh samples of stems,leaves,flowers and fruits of the two kinds of medicinal plants were harvested.Stereoscopy was used for the observation of macroscopic appearance of Ligustrum sinense Lour.and Jasminum elongatum (Bergium) Wild.,and the microscopy was used for the examination of their microscopic features of stem and leaf cross section,tear film and the powder of leaf upper and lower epidermis.Results Original plant characteristics of the two kinds of species were as follows:Ligustrum sinense Lour.had a racemes and funnel-shaped corolla,while Jasminum elongatum (Bergium)Wild.had a cymes and salverform corolla.Microscopic identification results were as follows:the stem pith of Ligustrum sinense Lour.was smaller,and the stem pith ofJasminum elongatum(Bergium) Wild.was bigger;Ligustrum sinense Lour.stem powder had less pit or texture in the sclereids,and had reticulate vessels,and Jasminum elongatum(Bergium) Wild.stem powder had apparent pit or texture in the sclereids,and had spiral vessels.Conclusion The results will provide a basis for the identification,exploitation and utilization of Ligustrum sinense Lour.and Jasminum elongatum(Bergium) Wild.

Objective To investigate the genetic polymorphisms of 27 Y-chromosomal short tandem repeats (Y-STR) loci included in Yfiler(R) Plus kit in Han population of Chaoshan area,and explore the population genetic relationships and evaluate its application value on forensic medicine.Methods By detecting 795 unrelated Chaoshan Han males with Yfiler(R) Plus kit,haplotype frequencies and population genetics parameters of the 27 Y-STR loci were statistically analyzed and compared with available data of other populations from different races and regions for analyzing the genetic distance and clustering relation of Chaoshan Han population.Results Seven hundred and eighty-seven different haplotypes were observed in 795 unrelated male individuals,of which 779 haplotypes were unique,and 8 haplotypes occurred twice.The haplotype diversity (HD) was 0.999975 with discriminative capacity (DC) of 98.99％.The gene diversity (GD) at the 27 Y-STR loci ranged from 0.3637(DYS391) to 0.9559(DYS385a/b).Comparing with Asian reference populations,the genetic distance (Rst) between Chaoshan Han and Guangdong Han was the smallest (0.0036),while it was relatively larger between Chaoshan Han and Gansu Tibetan population (0.0935).The multi-dimensional scaling (MDS) plot based on Rst values was similar to the results of clustering analysis.Conclusion Multiplex detection of the 27 Y-STR loci reveals a highly polymorphic genetic distribution in Chaoshan Han population,which demonstrates the important significance of Yfiler(R) Plus kit for establishing a Y-STR database,studying population genetics,and for good practice in forensic medicine.

Objective; To study the chromatic distribution of the chromatic value of Vitapan 3D- Master shade guide in color space. Methods: The color space of.Vitapan 3D-Master shade guide was measured and analyzed with CIE-1976-L* a* b. system and MINOLTA chromatic instrument (CR-321 ) under D65 standard source. Results: 26 shades of Vitapan 3D-Master could be divided into 5 groups according to the 3-dimensional layers. Inside the 2nd, 3th and 4th group, 6 shades distributed as equidistant ring circling M2 shade because their hue and chroma were different. The hue of L1. 5 and L2. 5 deflected to yellow, R1.5 and R2. 5 to red,that of M1-3 was intervenient. Conclusion: Vitapan 3D-Master shade guide is equidistant in color space and can be used more effeciently than Vitapan classical. Its chromatic value distribution perfectly and evenly covered color region of nature tooth.

Purpose To evaluate the feasibility of using dual-energy CT scanner to differentiate two contrast media bismuth and gadolinium. Materials and Methods Two phantoms containing contrast media of different ratio were scanned on dual-energy CT. Group 1 was mixture of iodine and gadolinium solution with volume ratio of 0∶1, 1∶6, 1∶1, 6∶1 and 1∶0. Group 2 was mixture of iodine and bismuth solution with mass ratio of 0∶1, 1∶6, 1∶1, 6∶1, 1∶0. Monoenergetic image reconstruction was performed at 80 keV. Liver VNC software was used for virtual scanning and iodine concentration analysis. Results Under 80 keV reconstruction, the measured CT attenuation of Group 1 was 379-383 HU, and 170-173 HU in group 2. The iodine concentration of two groups was not signiifcantly different between the calculated and actual iodine concentration (P>0.05). The dual-energy CT can distinguish two contrast media. Iodine and gadolinium contrast media were not statistically different, while iodine and bismuth contrast media agent could be easily differentiated. Conclusion Dual-energy CT can distinguish two contrast media with different attenuation.

Objective To investigate the effects of sulforaphane (SFN) on proliferation and invasion of human small cell lung cancer NCI-H446 and the activity of matrix metalloproteinase (MMP) -9.Methods NCI-H446 cells were cultured with 0,25,50,100 μmol/L SFN for 24 ～ 72 h,then MTT assay was employed to detect cell proliferation.Chamber invasion assay was used to study the cell invasion,and gelatin zymography assay was implied in MMP-9 enzyme activity.Results After treatment of 25,50,100μmol/L SFN,the growth of NCI-H446 cells were inhibited.When cells were incubated with 25,50,100μmol/L of SFN for 72h,the inhibition ratio was ( 11.1 ± 2.26 ) ％,( 25.2 ± 3.24 ) ％ and ( 44.6 ±4.2) ％,respectively,the difference was statistically significant compared with the solvent control group ( t =10.685,8.417,5.264,P ＜0.05 ).Chamber invasion assay showed that NCI-H446 cell invasion could be reduced.25,50,100 μmol/L of SFN could decrease the trans-membrane cells to (48.6 ± 1.84)％,(35.4 ± 2.22) ％ and (27.8 ± 1.36) ％,and it was statistically significant compared with the solvent control group ( t =6.341,5.562,4.925,P ＜0.05 ),respectively.In addition,MMP-9 activity was significantly inhibited by SFN.25,50,100 μmol/L of SFN could decrease the gray value of MMP-9 to 764 ±18.4,685 ± 14.74 and 638 ± 21.54 ( control group 822 ± 12.53,t =4.971,7.582,11.235,respectively,P ＜0.05).Conclusions SFN can inhibit NCI-H446 cells growth,invasion and the activity ofMMP-9.

Objective To evaluate the efficacy of CT in diagnosis of pulmonary blastoma (PB).Methods 10 patients with surgically and pathologically proved PB were collected in this study,and CT findings of the tumors were retrospectively analyzed. Results Of 10 cases,1 were central form (the tumors localized under pleura) and 9 was periphery form.The tumors were localized at right lobus in 6( including upper lobus,middle and lower lobus in 1,1 and 4,respectively) and left lobus in 4(upper lobus in 3 and lower lobus in one).The lesions were more than 3 cm in diameters in all cases.Necrosis of tumors were seen in 7 cases.The lesions had obvious enhancement after intravenous injection of contrast medium.Enlarged lymph nodes were identified at hilum of the lung and mediastinum in one,bony destruction and distant metastases were seen in one and 2,respectively.Conclusion PB is not of characteristic CT features,CT-guided percutaneous needle biopsy and the immunohistochemical method are helpful to the diagnosis of pulmonary blastoma.

OBJECTIVE: To observe the effects of Feiji Formula, a compound traditional Chinese herbal medicine, on lung cancer metastasis in mice. METHODS: The lung cancer metastasis model of mice was established in this experiment study. Twenty-four mice were randomly divided into three groups: untreated group, cisplatin group and Feiji Formula group. Mice in the Feiji Formula group were treated with Feiji Formula decoction; in cisplatin group, with cisplatin by intraperitoneal injection; and in the untreated group, with normal saline (NS). After twenty-day treatment, the body and tumor weights as well as the number of metastatic tumors in both lungs of each mouse were measured. RESULTS: The body weight of mice in cisplatin group was significantly less than that of Feiji Formula group and untreated group (P<0.01); the tumor weight of mice in cisplatin group and Feiji Formula group was markedly lower than that of untreated group (P<0.01); and the number of metastatic tumors in cisplatin group and Feiji Formula group was markedly lower than that of the untreated group (P<0.01), no significant difference between the Feiji Formula group and cisplatin group in terms of the weights and the numbers of metastatic tumors in bilateral lungs. CONCLUSION: Feiji Formula can suppress tumor growth and decrease the number of lung metastatic tumors in the mice, and maintain the body weight of the mice.

OBJECTIVE: To suppress NgR gene expression in neural stem cells and observe differentiation of neural stem cells in vitro after interfered which provide nutritional support for the facial nerve repair in vivo. METHOD: PCR amplification, restriction endonuclease digestion, T4DNA ligase connections were used to connected NgR with rector pGCsi, and constructed recombinant vector (NgR shRNA). Lipofectamine 2000 were used to transfect the NSC. The expression of NgR was examined by Western Blot. The proportion of neural stem cells transformed into neurons after transfection was tested by Immunocytochemistry. Neural stem cells were planted in PLGA tubes after transfected, and were scanned by electron microscopy. RESULT: NgR shRNA plasmid was constructed and infected neural stem cells successfully. Western Blot showed that the expression of NgR decreased in neural stem cells after interference. Immunocytochemistry showed that the rate of the neural stem cells transformed into neurons after interfered was significantly higher (P < 0.01). CONCLUSION Neural stem cells were transformed into neurons after NgR shRNA plasmid infected neural stem cells, which promoted axonal regeneration more effectively and provided a efficient and stable gene platform for facial nerve repair.
Animals ; Cell Differentiation ; Cells, Cultured ; Facial Nerve ; surgery ; GPI-Linked Proteins ; genetics ; metabolism ; Myelin Proteins ; genetics ; metabolism ; Neural Stem Cells ; cytology ; metabolism ; Nogo Receptor 1 ; RNA Interference ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; genetics ; metabolism