This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.
Actins ; genetics ; metabolism ; Animals ; Artemisia ; chemistry ; Artemisinins ; isolation & purification ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Epithelial Cells ; cytology ; metabolism ; Epithelial-Mesenchymal Transition ; drug effects ; Idiopathic Pulmonary Fibrosis ; pathology ; Plants, Medicinal ; chemistry ; Pulmonary Alveoli ; cytology ; RNA, Messenger ; metabolism ; Rats ; Smad3 Protein ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; pharmacology ; Vimentin ; genetics ; metabolism

Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

OBJECTIVESeizure is a common emergency in children with complicated pathogeny. Seizures are usually caused by complicated etiology and fever and febrile seizure are the commonest causes. Repeated and permanent seizures can damage the brain. So it is important to take active and effective measures to control seizure and high fever. Because most seizures and fever take place at home or out of hospital and it is difficult to administer drugs intravenously, it is important to explore an easy, safe, quick and effective way to control and prevent both seizure and fever. The present study aimed to explore the efficacy and safety of rectal administration of mixed ibuprofen and diazepam (IBU-DZP) solution.

METHODS(1) Animal study on the pharmacokinetics in rabbits and pharmacodynamics in rats after rectal administration with the mixed solution and on the irritability of the mixed solution to rectum. (2) Clinical study: Pharmacokinetics of the mixed solution in children after rectal administration were investigated.

RESULTS(1) Animal study: IBU and DZP were both rapidly absorbed from rectum with a peak blood level of (11.7 +/- 1.2) min and (9.4 +/- 2.7) min in rabbits, respectively. The mixed solution could effectively prevent the severity of seizures induced by pentetrazole and significantly suppressed fever induced by yeast. There were no remarkable pathological changes in rectal tissues after repeated rectal administration of the mixed solution. (2) Clinical study: IBU and DZP rapidly reached their peak blood levels at about 30 min and 15 min respectively after rectal administration to the children. The peak values were (57.8 +/- 7.9) mg/L and (450.1 +/- 158.7) microg/L, respectively. In fact, both of them reached levels that were much higher than their therapeutic levels in serum just at 5 min after administration, their blood levels were (41.4 +/- 5.5) mg/L and (321.8 +/- 53.9) microg/L, respectively.

CONCLUSIONSIBU-DZP mixed solution administered rectally is an easy, safe, quick and effective way to control and prevent both seizure and fever.

Administration, Rectal ; Animals ; Child ; Child, Preschool ; Diazepam ; administration & dosage ; pharmacokinetics ; Drug Therapy, Combination ; Humans ; Ibuprofen ; pharmacokinetics ; therapeutic use ; Infant ; Rabbits ; Rats ; Rats, Wistar ; Seizures, Febrile ; drug therapy

Objective To investigate the com and pepper fluorine in Zhenxiong county of Yunnan province, as well as the change of com and pepper fluorine after baked by coal, clay-mixed with cual for a relatively long-term, in order to provide a scientific basis for reducing fluorine intake. Methods The endemic areas of Yile, Wufeng's Songlinwan, Tangfang and Wufeng's Wugu in Zhenxiong county, and a non-endemic area Xiaguan in Dali city were selected as study sites. Ten samples of fresh corn and pepper were collected in each region, and fluorine was determined using acid leaching/potentiometry freshly and after baking or drying for 10 days or 4 months, respectively. Results The fluorine content of local fresh corn in Xiaguan of Dali city and Yile,Wufeng's Songlinwan, Tangfang, Wufeng's Wugu in Zhenxiong county were (1.31 ± 0.13),(1.65 ± 0.64),(1.92 ±0.37), (2.32 ± 0.49), (1.98 ± 0.66)mg/kg, respectively, and there were statistically significant differences across the regions(H = 27.871, P ＜ 0.05). The fluorine content of corn samples after baking or drying for 4 months were ( 1.82 ± 0.17), (26.43 ± 12.03), (39.27 ± 8.09), ( 14.27 ± 4.37), ( 14.33 ± 1.73)mg/kg, respectively, which were significantly higher than that of the fresh com in the corresponding region(all P ＜ 0.05 ), and there were statistically significant differences across the regions(H = 42.512, P＜ 0.05). The fluorine content of the local fresh chili were (3.34 ± 1.08), (3.44 ± 0.55), (3.47 ± 0.74), (3.46 ± 0.93)mg/kg, respectively, in the 4 observed places in Xiaguan of Dali city and Yile, Wufeng's Songlinwan, Tangfang in Zhenxiong county, and there were no statistically significant differences across the regions (F = 0.052, P ＞ 0.05 ). The fluorine content of pepper samples after baking or drying for 4 months were (7.01 ± 1.64), (226.07 ± 83.69), (179.36 ± 148.37), (54.51 ± 34.67)mg/kg,respectively, which were significantly higher than that of the fresh pepper in the corresponding region(all P ＜ 0.05 ),and there were statistically significant differences across the regions(H = 28.822, P ＜ 0.05). Conclusion Corn and chili fluorine is significantly increased after baked with coal and clay-mixed with coal by farmers in Zhenxiong county, a coal- burning borne endemic fluorosis area of Yunnan province.

Adult ; Aged ; Aged, 80 and over ; Female ; Hepatic Encephalopathy ; diagnosis ; Humans ; Male ; Middle Aged ; Neuropsychological Tests ; Psychometrics ; methods ; Young Adult

OBJECTIVETo evaluate the clinical stability of the palatal implant anchorage system in orthodontic treatment.

METHODSSixteen osseointegrated implants (5.0 mm in diameter, 6 mm length) were inserted in the median palatal suture area of 19 patients with malocclusion (average age: 18.22 +/- 7.10 years, from 11 years to 35 years 1 month) as anchorage of active orthodontic treatment with MBT appliance. The standard lateral cephalogram after implant placement and before implant removing was taken to compare the radiological parameters.

RESULTSThe successful rate of palatal implant anchorage was 84.2%. The average duration of 16 palatal implant was 23.08 +/- 8.06 months (from 10 months to 36 months). There were no statistical differences in all parameters from the implant placement to the end of treatment. IL-X was from (62.88 +/- 5.85) mm to (62.45 +/- 6.70) mm, IL-Y was from (36.66 +/- 5.41) mm to (37.96 +/- 4.90) mm, IAP-PP was from (73.81 +/- 8.84) degrees to (74.72 +/- 9.22) degrees, IAP-Y was from (62.09 +/- 9.33) degrees to (63.85 +/- 10.96) degrees, U6-Y is from (20.80 +/- 5.87) mm to (21.49 +/- 6.00) mm.

CONCLUSIONSStable palatal implant anchorage was maintained during active orthodontic treatment.

Adolescent ; Adult ; Child ; Female ; Humans ; Male ; Malocclusion ; therapy ; Orthodontic Anchorage Procedures ; instrumentation ; Orthodontic Appliance Design ; Palate, Hard ; surgery ; Young Adult

OBJECTIVEGastric cancer cells are insensitive to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). To sensitize gastric cancer cells to TRAIL, we treated gastric cancer MGC803 cells with TRAIL and cisplatin.

METHODSCell proliferation was measured using MTT assay. Cell apoptosis was determined by flow cytometry. Expression of proteins was analyzed by Western blot. The distribution of lipid rafts and death receptors was analyzed by immunofluorescence microscopy. MGC803 cells were pretreated with 50 mg/L nystatin for 1 h, and followed by the treatment of cisplatin and TRAIL.

RESULTS100 µg/L TRAIL resulted in (8.51 ± 3.45)% inhibition of cell proliferation and caused (3.26 ± 0.89)% cell apoptosis in MGC803 cells. Compared with the treatment with cisplatin alone, treatment with TRAIL (100 µg/L) and cisplatin (8.49 mg/L, IC(50) dose of 24 h) led to a dramatic increase in both inhibition of cell proliferation [(52.58 ± 4.57)% vs. (76.43 ± 5.35)%, P < 0.05] and cell apoptosis [(23.10 ± 3.41)% vs. (42.56 ± 4.11)%, P < 0.05]. Moreover, cleavage of caspase-8 and caspase-3 was detected. TRAIL (100 µg/L) did not induce obvious lipid rafts aggregation and death receptor 4 (DR4) clustering, while cisplatin (8.49 mg/L) significantly promoted the localization of DR4 in aggregated lipid rafts. Pretreatment with 50 mg/L nystatin, a cholesterol-sequestering agent, triggered (3.66 ± 0.52)% cell apoptosis after 24 h. Pretreatment with nystatin for 1 h before the addition of 8.49 mg/L cisplatin for 24 h caused a decreased tendency to cell apoptosis [(25.74 ± 3.28)% vs. (22.76 ± 2.97)%]. While, pretreatment with nystatin before the addition of cisplatin and TRAIL, the proportion of apoptotic cells decreased from (43.16 ± 4.26)% to (31.52 ± 3.99)% (P < 0.05).

CONCLUSIONCisplatin enhances TRAIL-induced apoptosis in gastric cancer MGC803 cells through clustering death receptors into lipid rafts.

Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Membrane Microdomains ; metabolism ; Nystatin ; pharmacology ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology

·AIM:To evaluate the efficacy of phacoemulsification and intraocular lens ( IOL ) implantation combined with goniosynechialysis in the treatment of primary chronic angle closure glaucoma.·METHODS: Eighty patients ( 96 eyes ) with primary chronic angle closure glaucoma were divided into observation group (40 cases, 46 eyes) and control group (40 cases, 50 eyes). The treatment group was treated with phacoemulsification, intraocular lens implantation and goniosynechialysis, while the control group was treated with phacoemulsification and intraocular lens implantation. The best corrected visual acuity, intraocular pressure, visual field mean deviation (MD), mean sensitivity ( MS) and the central anterior chamber depth of the two groups before and after the surgery were compared, and the occurrence of complications in patients with postoperative were record.·RESULTS:The best corrected visual acuity, visual field, intraocular pressure and central anterior chamber depth of the two groups before operation showed no significant differences (P>0. 05). The best corrected visual acuity, visual field, intraocular pressure and central anterior chamber depth between the groups at 6mo postoperatively showed statistically significant (P<0. 05). There was no serious complications in the two groups, and the difference between the two groups showed no statistically significant (x2=0. 351, P=2. 095).· CONCLUSION: The application of the phacoemulsification and intraocular lens implantation combined with goniosynechialysis in the treatment of primary chronic angle closure glaucoma is more effective than the simple use of phacoemulsification and intraocular lens implantation.

Objective To study the trend of accidental death among children under 5 years of age.Methods A survey on accidental death among children was performed in Guangzhou to compare the data from the national monitoring program on the deaths under five in the same period during 2001-2010.Data on mortality and causes of injures were adjusted by missing reports.Results Mortality rates on injuries were decreasing annually from 2001 to 2010.Comparing with 2001,the magnitude of the drop in mortality among the 0,1-4 and under 5 year groups were 72.02％,77.17％ and 74.66％ respectively,with statistically significant differences (P＝0.000).During the 10 years of observation,data showed a dropping trend (P＝0.000) of the mortalitics on drowning,traffic accident,suffocation and falling,especially on drowning and suffocation.The main causes of death were suffocation in the 0 year group,drowning in the 1-2-year group,traffic accident and poisoning in the 3-year group,traffic accident and falling in the 4-year group.The accidental deaths happened in ‘C area’ which including Huadu,Conghua,Zengcheng,Panyu and Nansha,with the total proportion of accidental death in Guangzhou as 75.69％.Conclusion The mortality of accidental deaths among children under 5 years of age in Guangzhou had been decreasing during 2001-2010,mainly owing to the practice of intervention programs on drowning and suffocation,suggesting the necessity of integrating the health education programs on accidental injury with the healthcare management system in children in the country,focusing on the suburban and county levels where economy was relatively undeveloped or with more floating population.

Global longitudinal strain (GLS) at rest on two-dimensional speckle tracking echocardiography (2D STE) was demonstrated to help detect coronary artery disease (CAD).However,the optimal cut-off point of GLS and its diagnostic power for detecting critical CAD in non-diabetes mellitus (DM) patients are unknown.In the present study,211 patients with suspected CAD were prospectively included,with DM patients excluded.All patients underwent echocardiography and subsequently coronary angiography within 3 days.Left ventricular (LV) GLSs were quantified by 2D STE.Territorial peak systolic longitudinal strains (TLSs) were calculated based on the perfusion territories of the 3-epicardial coronary arteries in a 17-segment LV model.Critical CAD was defined as an area stenosis ≥70％ in ≥1 epicardial coronary artery (≥50％ in left main coronary artery).Totally 145 patients were diagnosed as having critical CAD by coronary angiography.Significant differences were observed in all strain parameters between patients with and without critical CAD.The area under the receiver operating charcteristic (ROC) curve (AUC) for GLS in the detection of left main (LM) or threevessel CAD was 0.875 at a cut-off value of-19.05％ with sensitivity of 78.1％ and specificity of 72.7％,which increased to 0.926 after exclusion of apical segments (cut-off value-18.66％;sensitivity 84.4％ and specificity 81.8％).The values of TLSs were significantly lower in regions supplied by stenotic arteries than in those by non-stenotic arteries.The AUC for the TLSs to identify critical stenosis of left circumflex (LCX) artery,left anterior descending (LAD) artery and right coronary artery (RCA),in order of diagnostic accuracy,was 0.818 for LCX,0.764 for LAD and 0.723 for RCA,respectively.In conclusion,in non-DM patients with suspected CAD,GLS assessed by 2D STE is an excellent predictor for LM or three-vessel CAD with high diagnostic accuracy,and a higher cut-off point than reported before should be used.Excluding apical segments in the calculation of GLS can further improve the predictive accuracy of GLS.It is unsatisfactory for TLSs to be used to identify stenotic coronary arteries.