BACKGROUND: Icariin has a broad prospect for promoting cell proliferation. Differentiation direction of periosteal cells is uncertain, but the cells are easy to be induced by ultrasound, oxygen or bone morphogenetic protein 7 (BMP7). Periosteal cells have been applied in bone tissue engineering; however, icariin effects on the proliferation and differentiation of periosteal cells is little reported.OBJECTIVE: To investigate the effect of icariin on the proliferation and differentiation of human periosteal cells, thus providing theoretical basis for icariin applied in bone tissue engineering. METHODS:The human periosteum was obtained and the primary cells were isolated in vitro.After culture and expansion,periosteal cells were cultured in 24-well plates, and induced by 0.001, 0.01 and 0.1 mg/L icariin and 50 μg/L BMP7, respectively. The corresponding avsorbance values of different groups were detected. The levels of alkaline phosphatase and calcium nodules in periosteal cells were measured at 1, 3, 5 and 7 days, and the mRNA levels of osteocalcin, osteopontin and Runx2 were detected at 3, 5 and 7 days. RESULTS AND CONCLUSION: The periosteal cells proliferated well after induction with icariin, and could proliferate well in different concentrations of icariin and the positive control group (P < 0.05). Compared with the control group, the periosteal cells induced by icariin were able to produce more alkaline phosphatase and calcium nodules (P < 0.05). The mRNA expression of osteocalcin, osteopontin and Runx2 in periosteal cells could be up-regulated by icariin (P < 0.05). These findings imply that icariin can promote proliferation and differentiate of periosteal cells into osteoblasts, and it can be used as an inducer for the preparation of seed cells in bone tissue engineering.

BACKGROUND: Although icariin has a clear role in inducing differentiation of bone marrow stromal cells, no studies have reported on its use in proliferation of periosteal cells in animal experiments. Three-dimensional polycaprolactone scaffold is a new scaffold, and it has the characteristics of biodegradability, proper pore size, and good histocompatibility. OBJECTIVE: To investigate the effect of icariin on the proliferation of periosteal cells, to construct periosteal cells and three-dimensional scaffold composite tissue-engineered bone and study its therapeutic effect on radical defects in rabbits, so as to provide a new way for the treatment of bone defects . METHODS: The mandibular periosteum was obtained from the rabbit and the primary cells were obtained by tissue explant method. The passage 3 cells were planted on 24-well plates. In experimental group, icariin at 10-4, 10-3and 10-2mol/L were added into the corresponding cell culture wells; equal volume of PBS was added to control group; positive control group was treated with 10 μg/L basic fibroblast growth factor. The cell proliferation in each group was measured by MTT method on the 2nd, 4th, 6thand 8thdays, respectively. The three-dimensional polycaprolactone scaffolds,10-2mol/L icariin and 107/L periosteum cells were placed in cell culture plates and cultured in vitro to construct the cell-scaffold complex. After the rabbit radical defect model was established, the model rabbits were randomized into three groups (n=15 per group), followed by implanted with scaffold complex (experimental group), polycaprolactone scaffold (polycaprolactone group) or nothing (control group). Pathological sections were taken at the 2nd, 4thand 8thweeks respectively for hematoxylin-eosin staining to observe the bone and count the number of osteoblasts. The X-ray films of the radius were obtained at the 4thweek. RESULTS AND CONCLUSION: (1) The proliferation of periosteal cells induced by different concentrations of icariin, especially 10-2mol/L icariin, was better than that in the control group (P < 0.05), Therefore, the optimal concentration of icariin was 10-2mol/L. (2) At 4 weeks after transplantation, in the experimental group, there were more osterocytes and less microvessels; in the polycaprolactone group, there were more new vessels, less woven newly born bone and osterocytes; in the control group, the defect region was filled with more granulation tissue and less osterocytes. At the 8thweek, complete healing in the defect region was observed in the cell-scaffold complex group, while partial healing in the polycaprolactone scaffold group. In the control group, however, fibroblasts and scar tissues were visible in the defect region, with presence of closed medullary cavity and nonunion. The number of osteocytes in the experimental group was significantly higher than that in the other two groups (P < 0.05). X-ray examination at the 4thweek showed that the defect region in the experimental group achieved healing;in the polycaprolactone group, the callus formed and defect region connected partly; and there was a clear gap in the defect region of the control group. Our findings indicate that icariin at different concentrations can promote the proliferation of periosteal cells and the concentration of 10-2mol/L has the greatest role. Icariin can induce proliferation of periosteal cells in the three-dimensional polycaprolactone scaffold, which contributes to bone repair.

Objective To study the changes of soluble intercelluar adhesion molecule-1 (sICAM-1) in serum of neonates with hy-poxic-ischemic encephalopathy (HIE),and significance of changes of serum sICAM-1 in HIE pathogenesis. Methods There were 17 controls of neonates and their sICAM-1 concentrations in serum were detected with enzyme-linked immunosorbent assay (ELISA) at the critical stage and at the beginning of convalescent stage in 36 cases of HIE neonates and 17 normal neonates. The data were analyzed with analysis of variance, Newman-Keuls q test. Results The concentrations of sICAM-l[(216.64?85.32)?g/L] at the critical stage of 36 cases HIE neonates were significantly higher than those [(6. 16?4.05) ?g/L ] of control group (q = 17. 42 P

Objective To investigate the effect of prolonged stroboscopic illumination exposure on the growth of eyeball of guinea pig. Methods Thirty 2-week-old guinea pigs were randomized into three groups ( n=10 for each) . Two strobe-reared groups were raised with 20 Hz sinusoidal and 20 Hz square wave stroboscopic illumination, respectively. The control group received usual light illumination. The illumination intensity was 500 lux. All animals underwent refraction and biometric measurements prior to and after 2, 4, 6 and 8 weeks of treatment. Finally, flash electroretinograms were com-pared, and retinal microstructures were examined. Results There was a significant correlation between refractive errors and axial eye elongation, and myopia increasing was observed with eye elongation. After 8 weeks of treatment, the animals raised in 20 Hz sinusoidal and 20 Hz square wave stroboscopic illumination were (-0. 75 ± 0. 79)D and (-1. 50 ± 0. 91) D more myopic than the group raised in continuous illumination. The implicit time of the a-wave was delayed by 3. 8 and 7. 9 ms, respectively. No significant difference was found in retinal ultrastructures among the three groups. Conclusions Chronic exposure to 20 Hz sinusoidal or square wave stroboscopic illumination alters the emmetropization of the guinea pig eye to some extent.

Objective: To investigate the function and mechanism of phospholipase Cγ1 (PLCγ1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-κB) were used to study the effect of PLCγ1 and NF-κB on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLCγ1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLCγ1 or NF-κB resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P<0.05), but had no marked effect on SW480 cells. Western blot analysis showed that epidermal growth factor (EGF) stimulated the phosphorylation of PLCγ1 in LoVo. The results of EMSA indicated that inhibition of PLCγ1 signaling pathway also down-regulated the activity of NF-κB while EGF reversed the function. Conclusion:These data suggest that PLCγ1 plays a pivotal role in the EGF-induced cell-matrix adhesion of highly metastatic colorectal cancer cells and that NF-κB is also functional in this signaling pathway.

Objective To investigate the prevalence of diabetes and impaired glucose regulation in high risks community residents in Songjiang District of Shanghai, and to explore the risk factors affecting their blood glucose metabolism, providing effective suggestions for improving community diabetes prevention and treatment. Methods Questionnaire, physical examination and laboratory testing were used to collect information on the basic characteristics and blood glucose levels of 21 035 residents in Songjiang District who were assessed to be at high risk of diabetes. Results A total of 3 008 people with impaired regulation and 2 241 patients with diabetes were detected.The detection rates were 14.3% and 10.6%, respectively.It was found that as the residents′ age was higher, their education level was lower, high-risk factors for them were numerous, and their detection rates of diabetes and impaired sugar regulation were higher.It was found that age, gender, history of impaired glucose regulation, relatives with type 2 diabetes, hypertension, dyslipidemia and overweight/obesity were all factors influencing blood glucose metabolism (P < 0.05). Conclusion Blood glucose screening in high-risk population of diabetes is helpful for the early detection of diabetic patients and those with impaired glucose regulation.It is necessary to strengthen the attention of population at risk of diabetes and take appropriate interventions.

OBJECTIVETo compare the clinical efficacy in the treatment of allergic rhinitis (AR) of lung qi deficiency and cold syndrome between Jin's three-needle therapy and western medication.

METHODSSixty-six patients were randomized into an acupuncture group and a western medication group, 33 cases in each one. In the acupuncture group, acupuncture was applied at three-nose points [Yingxiang (LI 20), Shangyingxiang (EX-HN 8) and Yintang (GV 29); Cuanzhu (BL 2) was added for frontal headache] and three-back points [Dazhu (BL 11), Fengmen (BL 12) and Feishu (BL 13)], once every day. Ten treatments made one session. Two sessions of treatment were required. In the western medication group, desloratadine oral suspension was prescribed, 5 mg each time, once a day, for 20 days. The scores of the symptoms and physical signs in AR patients as well as the clinical efficacy were observed between the two groups.

RESULTSThe total effective rate was 93.9% (31/33) in the acupuncture group, which was better than 72.7% (24/33) in the western medication group (P < 0.05). After treatment, the scores of AR symptoms and physical signs as well as the total score were all reduced compared with those before treatment in the two groups (all P < 0.01). The score of every item in the acupuncture group was lower than that in the western medication group after treatment (score of symptoms: 4.70 +/- 2.07 vs 6.55 +/- 2. 69, score of physical signs: 0.85 +/- 0.67 vs 1.45 +/- +0.62, total score: 5.36 +/- 2.70 vs 8.00 +/- 2.91, all P < 0.01).

CONCLUSIONJin's three-needle therapy achieves superior efficacy on AR of lung-qi deficiency and cold syndrome, which is better than desloratadine oral suspension.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adolescent ; Adult ; Child ; Female ; Humans ; Lung ; physiopathology ; Male ; Middle Aged ; Needles ; Qi ; Rhinitis, Allergic ; Rhinitis, Allergic, Perennial ; physiopathology ; therapy ; Young Adult

Squalene synthase (SQS) is a key enzyme in plant terpenoid biosynthetic pathway. This study focused on cloning and analysis of Huperzia serrata SQS (HsSQS1) gene. After searching the transcriptome dataset of H serrata, one unique sequence encoding SQS was discovered. The primers were designed according to the transcript sequence of HsSQS1 from the H. serrata transcriptome dataset. The open reading frame of HsSQS1 was cloned using RT-PCR strategy. The bioinformatic analysis of this gene and its corresponding protein were performed. The cDNA (named as HsSQS1) contains a 1263 bp open reading frame and encodes a predicted protein of 420 amino acids. The GenBank accession number for this gene is JQ004938. HsSQS1 contains two transmembrane regions, without signal peptide. The conserved domain of squalene synthase was presented in HsSQS1. HsSQS1 was more abundant in H. serrata root than in leaf and stem. This study cloned and analyzed squalene synthase gene from H. serrata for the first time. The result will provide a foundation for exploring the mechanism ofterpenoid biosynthesis in H. serrata plants.
Amino Acid Sequence ; Biosynthetic Pathways ; Cloning, Molecular ; DNA, Complementary ; genetics ; Expressed Sequence Tags ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; isolation & purification ; metabolism ; Genes, Plant ; genetics ; Huperzia ; enzymology ; genetics ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Plant Stems ; enzymology ; Plants, Medicinal ; enzymology ; genetics ; Triterpenes ; chemistry

Objective:To improve the genetic stability of HBV gene in transgenic mice.Methods:HBV transgenic mice were bred by backcross and double cross.The HBV gene expression and replication were studied with real-time PCR,ELISA and chemiluminescence.Results:The HBV transgenic mice have stably bred to 23rd generation.The serum HBsAg level is 4122.31?2044.74IU/ml;The rate of HBV transgenic mice whose serum HBV DNA reach 104~106copies/ml was 93.93%.The HBV replication and expression were improved markedly.There is no difference between male and female mice about serum HBsAg level.Conclusion:After breeding the HBV gene was expressed stably with high-level in transgenic mice.

OBJECTIVETo explore the causes of nonspecific chronic cough in children and relationship between transient receptor potential vanilloid 1 (TRPV1) gene polymorphisms and nonspecific chronic cough.

METHODSA total of 195 children with chronic cough were followed up half a month, one month and three months after their first visit to hospital. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to examine polymorphisms of the TRPV1 gene in the children. A total of 205 healthy or surgical children without chronic cough served as the control group.

RESULTSThe etiologic distribution of the 195 children with chronic cough was as follows: 96 (49.2%) cases of cough variant asthma (CVA), 48 (24.6%) cases of CVA complicated by upper airway cough syndrome (UACS), 34 (17.4%) cases of post-infectious cough, and 17 (8.7%) cases of UACS. Three genotypes were identified in both groups at positions rs222747 (CC, GC and GG), rs222748 (CC, TC and TT) and rs8065080 (CC, TC and TT). The frequencies of genotype and allele at position rs222747 did not accord with the law of Hardy-Weinberg. There was no significant difference in frequencies of genotype and allele at positions rs222748 and rs8065080 between the two groups.

CONCLUSIONSCVA, UACS and post-infectious cough are common causes of nonspecific chronic cough in children. TRPV1 gene polymorphisms at positions rs222748 and rs8065080 may be unrelated to nonspecific chronic cough in children.

Adolescent ; Alleles ; Child ; Child, Preschool ; Chronic Disease ; Cough ; etiology ; genetics ; Female ; Genotype ; Humans ; Infant ; Male ; Polymorphism, Genetic ; TRPV Cation Channels ; genetics