Objective To observe the effects of epidermal growth factor (EGF) on cell proliferation and cell cycle of cultured bovine corneal endothelial cells. Methods Bovine corneal endothelial cells were cultured with different concentrations of hEGF (1, 10 and 100 ng/mL). MTT test was performed to evaluate the cell proliferation. The bovine corneal endothelial cells were divided into two groups:control group (cultured in DMEM) and EGF-stimulated group (cultured in DMEM with EGF). Flow cytometry was performed to determine the cell cycle phases on the third and seventh day. Results Compared with the control, EGF enhanced the cell proliferation in a dose-related response. 10 ng/mL and 100 ng/mL EGF were much more effective than 1 ng/mL.On the third day, S phase cells accounted for 24.5% and G_2-M phase cells 0.08% in the control group,while 24.6% and 0.06%, respectively in the EGF-stimulated group. However, on the seventh day, those came to 20.8% and 0.41% in the control group, and 18.2% and 1.55% in the EGF-stimulated group,indicating a significant change in the cell cycle (P

3T3 Cells ; Animals ; Cell Membrane ; metabolism ; Mice ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; radiation effects ; Radiation, Ionizing ; Wound Healing ; radiation effects

In order to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix using ITS2 barcodes, genomic DNA from sixty samples was extracted and the ITS2 (internal transcribed spacer) regions were amplified and sequenced. The genetic distances were computed using MEGA 5.0 in accordance with the kimura 2-parameter (K2P) model and the neighbor-joining (NJ) phylogenetic tree was constructed. The results indicated that for Aucklandiae Radix (Aucklandia lappa), Vladimiriae Radix (Vladimiria souliei and V. souliei var. cinerea), Inulae Radix (Inula helenium), Aristolochiae Radix (Aristolochia debilis) and Kadsurae Radix (Kadsura longipedunculata), the intra-specific variation was smaller than inter-specific one. There are 162 variable sites among 272 bp after alignment of all ITS2 sequence haplotypes. For each species, the intra-specific genetic distances were also smaller than inter-specific one. Furthermore, the NJ tree strongly supported that Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix can be differentiated. At the same time, V. souliei (Dolomiaea souliei) and V. souliei var. cinerea( D. souliei var. cinerea) belonging to Vladimiriae Radix were clearly identified. In conclusion, ITS2 barcode could be used to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix. Our study may provide a scientific foundation for clinical safe use of the traditional Chinese medicines.
Aristolochia ; classification ; genetics ; Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Quality Control

Objective To explore the expression of tankyrase (TNKS) and its relationship with WNT/β-catenin signal?ing pathway in lung acinar adenocarcinoma. Methods Seventy-two samples of single subtype alveolar like lung adenocarci?noma (lung adenocarcinoma group) and 67 specimens of normal lung tissue adjacent to carcinoma (adjacent to carcinoma group) were collected. Immunohistochemical method was used to detect expressions of TNKS, beta-catenin (β-catenin) and c-myc protein. The correlation of each protein expression in lung adenocarcinoma tissues was analyzed. The differential ex?pression of TNKS was detected by Western blot assay in two groups. Results Tankyrase protein was mainly expressed in cy?toplasm. The expression ofβ-catenin protein was mainly in cytoplasm and nuclear of lung adenocarcinoma. The expression ofβ-catenin was mainly in cytoplasm, and a small amount was in nuclear of the adjacent group. The c-myc protein was ex?pressed mainly in the nucleus. The positive expression rates of TNKS,β-catenin and c-myc protein were significantly high?er in lung adenocarcinoma group than those of adjacent to carcinoma group (P<0.05). The expression ofβ-catenin in cyto?plasm and nucleus was positively correlated with the expression of TNKS and c-myc (P<0.05). Western blot analysis showed that the relative expression level of TNKS was significantly higher in lung adenocarcinoma group than that of adja?cent to carcinoma group (0.497 ± 0.021 vs. 0.237 ± 0.015, t=13.00, P<0.01). Conclusion Abnormally high expression of TNKS in lung adenocarcinoma may promote the occurrence of lung cancer by regulating the WNT signaling pathways. Inhib?iting TNKS expression may become a new target to treat lung adenocarcinoma.

The identiifcation of body lfuid and their tissue resource is an important part of forensic medicine research. Conventional methods have imperfections like high false positive rate and may destroy biomaterial, which should be replaced by a new conifrmatory test. Highly-differentiated cells express speciifc mRNA molecules that can be used for body lfuid identiifcation. These mRNA markers can identify body lfuids like peripheral blood, menstrual blood, semen, saliva, vaginal secretion, skin-contact stains. This method have favorable sensitivity, speciifcity and can be detected after years, made it a ideal way to identify current body lfuids in the future. This review focus on the application of mRNA in body lfuid identiifcation and tissue resource.

ObjectiveTo study effects of oral administering alfa calcidol and diphosphonates on the bone mineral density (BMD) of patients with spinal cord injury (SCI).MethodsChanges of BMD of the 4th lumbar spine (L4), the proximal femur (total) and all of its subareas were studied between SCI cases of taking (n=31) and not taking (n=42) alfa calcidol. Changes of BMD between cases of taking diphosphonates (n=22) and not taking (n=24) were also compared.ResultsThere were no significant differences in BMD of L4, the proximal femur and all of its subareas between SCI cases of taking and not taking alfa calcidol. While there was a significant difference in BMD of the proximal femur (total) between cases of taking and not taking diphosphonates (P<0.05), and the reducing degree of BMD in the former is less than that in the latter.Conclusion Oral administering diphosphonates has a protective effect on SCI patients' BMD of the proximal femur.

OBJECTIVETo investigate the effects of occlusal rehabilitation on chewing patterns of patients with extensive tooth wear.

METHODS29 patients with severe tooth wear were selected and divided into two groups: Group I with complete posterior tooth support (15 cases) and group II with one-side or both side posterior support lost (14 cases). 15 normal old persons were also selected as control group. The surface electromyography (EMG) of masseter(MM), anterior temporalis (TA) and posterior temporalis (TP) during chewing movement were recorded in the stage of pre-treatment, 1 month after temporary restoration and 1 month after permanent restoration. The EMG activity, total cycle duration (TCD) and ratio of activation period to relaxation period (AP/RP) of chewing cycles were measured and compared.

RESULTS1) Before treatment, the TCD of these two groups were longer than normal group, but the differences were not significant (P > 0.05). All of the AP/RPs of MM, TA and TP in group II were significantly higher than that of normal group (P < 0.01), and EMG activity of TA of group II was group I were both significantly lower than that in pre-treatment stage. 3)After permanent restoration, the TCD of group I were significantly lower than that in temporary restoration stage (P < 0.01). In group II, the TCD was continuously slightly shortened, while the AP/RP were significantly lower (P < 0.01) and EMG activities were significantly higher(P < 0.01) than that in the temporary restoration stage.

CONCLUSIONOcclusal rehabilitation could obviously change the chewing patterns and the EMG activities of patients with severe tooth wear.

To investigate the expression patterns of peroxisome proliferator activated receptor2 (PPARgamma2) gene in the differentiation of mouse embryonic stem (ES) cells into adipocytes, mouse ES cells were transfected with the vector of pPPARgamma2-promoter-luciferase, and PPARgamma2 expressions were analyzed by detecting luciferase activities and by detecting the protein expressions using western blotting. The results showed that the gene PPARgamma2 did not express in undifferentiated mouse ES cells and in embryoid bodies (EBs) within the first 2 days of differentiation induction after EB formation, and began to express from the third day of differentiation induction after EB formation to the finish of the differentiation. The gene's expression in differentiated adipocytes was much stronger than that in differentiating preadipocytes. In Conclusion our results reported for the first time the five-step expression patterns of the gene PPARgamma2 during the whole differentiation procedures from mouse ES cells into adipocytes via preadipocytes, and supported the previous studies that PPARgamma2 is a fat-specific gene that expresses only in developed and developing adipose tissues.
Adipocytes ; cytology ; Animals ; Cell Differentiation ; genetics ; Cells, Cultured ; Electroporation ; Embryonic Stem Cells ; cytology ; metabolism ; Mice ; PPAR gamma ; biosynthesis ; genetics ; Promoter Regions, Genetic ; genetics ; Transfection

A cell model is desired for adipocyte differentiation investigation and for high-throughput screening of anti-obesity and anti-diabetes molecules from chemical resources due to the world wide epidemic of obesity and diabetes. In order to establish such a cell model, a plasmid of pPPARgamma2-promoter-EGFP was constructed by inserting a 660bp sequence of mouse PPARgamma2 promoter into the Ase I and Kpn I sites of pEGFP-N3 and transferred into 3T3-L1 preadipocyte cells. The cells were induced to differentiate and the expression of PPARgamma2 was detected by the microscopic observation of EGFP and by RT-PCR assays. The results showed that the EGFP gene expression patterns were similar to that of pPPARgamma2's, which indicated that the EGFP gene was transferred into the mouse 3T3-L1 preadipocyte cells, and its expression was under the control of pPPARgamma2 promoter. RT-PCR assays showed that the EGFP expression authentically represented the stable expression of PPARgamma2. In conclusion, a preadipocyte cell line expressing EGFP under the control of the promoter of adipocyte-specific expression gene PPARgamma2 was generated. The cell line provides a powerful approach for the research of adipocyte differentiation and for the high-throughput screening of anti-obesity and anti-diabetes chemicals.
3T3-L1 Cells ; Adipocytes ; cytology ; Animals ; Cell Differentiation ; Green Fluorescent Proteins ; genetics ; Mice ; PPAR gamma ; genetics ; Promoter Regions, Genetic ; Stem Cells ; cytology

Comrade ZHU Lian is a main founder of Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences, and she is the first to learn TCM for a physician of western medicine. The present paper looks back the works of comrade ZHU in acupuncture and moxibustion cause. Her main contributions are learning acupuncture and moxibustion medicine, popularizing acupuncture and moxibustion therapy; being concerned acupuncture and moxibustion cause, establishing organization of acupuncture and moxibustion research, more early conducting international exchanges of acupuncture and moxibustion; paying attention to clinical practice, proposing scientific research of acupuncture and moxibustion; writing New Acupuncture and Moxibustion Science, passing on later generations.
Acupuncture ; China ; History, 20th Century