Objective To construct an artificial skin model of melanoma by mixed culture of human keratinocytes and MV3 melanoma cells on de-epidermized dermis (DED) in order to study the effect of keratinocytes on melanoma invasion. Methods Epidermal cell suspension was obtained by a two-step digestion method from the circumcised foreskin of a child, keratinocyte serum-free medium was applied to the culture and passage of keratinocytes. MV3 melanoma cells were cultured and passaged in RPMI 1640 medium. Log-phase keratinocytes and MV3 cells were mixed with a ratio of 3:1 and seeded onto the surface of DED followed by a liquid culture and air-liquid culture for a total of 2 weeks. Thereafter, the artificial tissue model was assessed by HE staining and immunohistochemical staining for S-100 protein, HM64S and keratin. Results HE staining showed that MV3 cells formed band-like tumor masses or foci on the surface of DED, with keratinocytes intermingling among the tumor cells, but no typical epidermis-like structure was observed. Some tumor cells infiltrated into the surface of DED and showed a cluster distribution; some tumor cells invaded the lumen of the DED, and attached to the luminal wall in a ring shape; some tumor cells penetrated through the wall into the surrounding dermal tissue. On the bottom and lateral side of DED, tumor cells were infiltrating dispersedly.The tumor loci stained positive for S-100 protein and keratin, and weakly positive for HMB45. Conclusion Keratinocytes enhance the invasion of MV3 melanoma cells into the skin tissue model of melanoma.

Objective To investigate effects of combined use of glycogen synthase kinase (GSK) -3 inhibitor and fructose-1,6-diphosphate (FDP) on liver trauma in rats. Methods After crea-tion of liver trauma model in 49 Sprague-Dawley rats, 42 rats were randomly divided into control group (NaCl group), FDP group and FGI Group (FDP and GSK-3 inhibitor in combination group). Then, each group was randomly subdivided into pre-ischemia group and 4-hour reperfusion group on account of time point when animals were sacrificed before and after iachemia. The other seven rats set as sham operation group (SH group) were sacrificed at 4-hour reperfusion time point. The AST and ALT levels in hlood and glycogen content, SOD vitality and MDA content in liver tissues were determined. Results At pre-is-chemia time point, liver glycogen content in three groups was in order of control group < FDP group < FGI group (P <0.01). At 4-hour reperfusion time point, blood ALT and AST levels in four groups were in order of control group > FDP group > FGI group > SH group (P < 0.01), while SOD vitality in liver tissues of four groups was in order of control group < FDP group < FGI group < SH group (P < 0.01) and MDA content in four groups was in order of control group > FDP group > FGI group > SH group (P < 0.01). Conclusions Combined use of FDP and GSK-3 inhibitor can enhance the protective effect of FDP on liver rupture, as may relate to the mechanism that GSK-3 inhibitor can effectively enhance glycogen synthesis of FDP as substrate before liver ischemia so that the liver glycogen storage is increased in a short period of time and hence post-traumatic warm ischemia-reperfusion injury is alleviated in the liver of rats.

Objective To evaluate the effect of oral health instructions in nursing of marsupialized jaw cysts. Methods 46 patients with cysts of jaw were divided randomly into two groups (education group and control), 23 in each. In addition of normal nursing processes in both groups, patients in education group were offered more oral health instructions, such as pathogenesis of jaw cysts, mechanism of marsupialization, wear and clean of a cyst plug, and oral hygiene maintenance. 12 months later, compliance, satisfaction, treatment effect and oral hygiene condition of the patients in the two groups were studied and compared. Results 100%of patients in the education group could return to clinical visits on time and 96%of patients did cyst rinsing after every meal, which were significantly higher (χ2=6.9, P<0.05) than in the control (74% and 70% respectively). Patients′satisfaction in the two groups were also significantly different (χ2=9.109,P<0.05). Patients in the education group were much more satisfied with the treatment procedures than control. The treatment effect of the cyst in the education group [good (83%), moderate (17%), not so good (0%)] was significantly (χ2=7.793, P<0.05) better than control (good (43%), moderate (52%), not so good (5%)]. The secondary infection rates in the two groups were similar (χ2=1.022, P>0.05). Oral hygiene condition of patients in the education group was better than control in Debris Index (χ2=9.576, P < 0.05) and Plaque Index (χ2=8.212, P < 0.05). Conclusions Oral health instructions played positive role in improving patients′ compliance, degree of satisfaction, treatment effect and oral hygiene condition in patients with jaw cysts.

Objective To explore the apoptosis of K562/G01 cells induced by triptolide through MDM2/p53 signaling pathway. Methods K562/G01 cell line was treated with different concentrations of triptolide. MTT was used to detect the cell proliferation inhibition rate. FCM was used to determine the apoptosis rate changes in 12 h and 24 h. The mRNA expression levels of bcr-abl, XIAP, MDM2, p53 were detected by real-time quantitative PCR. Results After treatment by 10, 20, 40, 80, 100 nmol/L TP in 12, 24, 48 h, the viability of K562/G01 cells was inhibited in time-dose dependence manner. K562/G01 cells was treated by 20 nmol/L, 40 nmol/L TP after 12 h, 24 h, the cell apoptosis rate was rising with drug concentration and time. The bcr-abl, XIAP, MDM2 mRNA expression was down-regulated and p53 mRNA expression was up-regulated by TP. Conclusion TP can inhibit the growth of K562/G01 cell line and induce apoptosis through XIAP-MDM2-p53 signaling pathway.

Objective To investigate the effect of arsenic(As2O3) on spermatogenic cell apoptosis and the expression of telomerase activity in adult rats.Methods 40 healthy male Sprague-Dawle rats were randomly divided into four groups and they were treated with As2O3 at doses of 0(control group),0.375,0.75 and 1.5 mg/kg body weight respectively through gavage for 16 consecutive weeks.The numbers of testicular sperm head were counted and the coefficient of testicular viscera,daily sperm production(DSP) were calculated in every group.The apoptosis of germ cell was assessed by in situ terminal deoxynucleotityl transferase mediated dTUP nick end labeling(TUNEL) technique.The activity of telomerase in the spermatogenic cells was determined by immunohistochemistry.Results In the testes of As2O3-treated rats,the coefficient of testicular viscera,the number of testicular sperm head and DSP in moderate and high dose groups decreased significantly than that of control(P

0.05), but the patients in the rehabilitation group demonstrated much higher scores in Fugl Meyer and Barthel index than those in the control group at the end of 4 and 12 weeks after rehabilitation therapy( P 0.05) 12 weeks after the therapy. Conclusion Early intensive rehabilitation therapy can effectively improve the motor function and ADL of stroke patients; the continuing home rehabilitation program for at least 8 weeks is helpful for improving the patient's functional independence, increasing the cost effectiveness, and shortening the length of stay of stroke patients.

Objective To evaluate the accuracy and comparability of secondary hematology analyzer for platelet enumeration in order to determine the accuracy and reliability of assigned value of fresh blood.Methods The results between secondary standard hematology analyzer and the reference method of platelet enumeration of 40 specimens were compared according to the document from CLSI EP9-A2.The correlation and bias were calculated.At the same time,the results of secondary standard hematology analyzer between our laboratory and Japan reference laboratory were compared.The fresh blood from normal people was prepared to be used as calibrator after assigned value by secondary standard hematology analyzer.And 36 hematology analyzers were performed correctness validation and calibrated by 36 fresh bloods.Results The results of 40 specimens by secondary standard hematology analyzer and the reference method were ( 108 -326) × 109/L and( 110 -327 ) × 109/L respectively.Correlation coefficient between the secondary standard hematology analyzer and the reference method was 0.993.The bias between two methods was from -3.8％to 3.4％.The results of NCCL and Japan reference laboratory from 2009 to 2010 were( 185 -203) × 109/L and (185 - 198) × 109/L The bias range between our laboratory and reference laboratory in Japan was from - 1.4％ to 3.7％.The ranges of coefficient variations of two laboratories were from 2.0％ to 3.0％ and from 2.6％ to 3.4％,respectively.The biases of 20 hematology analyzers were from - 2.6％ to 2.1％ and they passed the correctness validation.The biases of 16 hematology analyzers were decreased from 3.4％ - 12.6％of pre-calibration to 0％ - 2.8％ of post-calibration.Conclusions The results of secondary standard hematology analyzer are assured to be accurate and comparable by the comparison of reference laboratories.It is feasible that fresh blood assigned value by secondary standard hematology analyzer can be used as calibrator for the hematology analyzer.

A reusable chronocoulometric adenosine triphosphate (ATP)-aptamer sensor was developed in this work.A short chain of DNA marked as cDNA containing complementary sequence was immobilized on gold electrode based on Au-S self-assembly.The ATP aptamer was hybridized with cDNA.The surface-confined DNA could bind with [Ru(NH3)63+ (RuHex) in the electrolyte via electrostatic interaction.Upon target ATP binding, the aptamer confined onto electrode surface was disassociated from the cDNA oligonucleotides into the solution.Such surface density change of DNA lead to the decrease of chronocoulometric signal for the RuHex which confined on the electrode surface.The chronocoulometric signals showed a linear relationship with logrithm of ATP concentration in the range of 1 nmol/L to 100 μmol/L, and the detection limit of this aptamer sensor could reach 0.5 nmol/L (S/N=3).This aptamer sensor could be regenerated 5 times by simple steps.With this aptamer sensor, the basal level of ATP in the brain cortex micorodialysate was determined to be 19.2±3.7 nmol/L (n=3).

ObjectiveTo explore the effects of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor-kappa B (NF-κB) signal pathway on the process of follicle-stimulating hormone (FSH) facilitating cell proliferation and invasion in human epithelial ovarian cancer. Methods Ovarian cancer cell lines SKOV3 and 3AO were cultured to exponential phase,then assigned to control group,FSH group,LY294002 group and FSH + LY294002 group,respectively.Cells were treated with different concentration of FSH and LY294002,respectively.The effects of FSH on cell proliferation were observed by methylthiazolyl tetrazolium (MTT).Morphological changes were observed by phase contrast microscope.The ability of cell invasion was investigated by transwell invasion assay.The expression of FSH receptor (FSHR),Akt1/2,phosphorylated-Akt (p-Akt) and NF-κB p65 protein were detected by western blot.Results( 1 ) FSH could promote the proliferation of SKOV3 and 3AO cells.When the cells were treated with 40 U/L FSH for 48 hours (SKOV3) and 24 hours (3AO),compared with those in control groups,they reached the highest proliferation rate (P ＜ 0.05 ),respectively.(2) The morphology of SKOV3 and 3AO cells in four groups:in control group,SKOV3 cells were short spindle and 3AO cells were long spindle,the nuclei of them were both roundness or oval,the cytoplasm were bright.In FSH group,the cells changed to slightly longer or polygonal,they were full in shape,meanwhile,the cell intensity were higher than control group.In LY294002 group,some cells changed from spindle to round,and began to shrink.The cell intensity diminished.The morphology of FSH + LY294002 group was similar with control group,but the cell intensity was lower than that in FSH group.(3)The number of SKOV3 cell that passed through the membrane in control group,FSH group,LY294002 group and FSH + LY294002 group was (26 ± 6),( 118 ± 19),( 18 ± 5) and ( 38 ± 7 ),respectively.The number of 3AO cell was ( 19 ± 4 ),( 134 ± 20),(12 ±3) and (58 ± 11 ),respectively.The results showed that the number of cells in FSH group was significantly higher than that in control group ( P ＜ 0.05 ),while the number of cell in FSH + LY294002 group was significantly fewer than that in FSH group (P ＜ 0.05 ).(4) There was no significant difference in the expression of FSHR and Akt1/2 between FSH group and control group (P ＞ 0.05 ),but FSH increased the expression of p-Akt and the ratio of NF-κB p65 in the nucleus versus cytoplasm in SKOV3 and 3AO cells,there were significant differences compared with control group ( P ＜ 0.05 ).LY294002 reversed the effects of FSH on increasing the expression of p-Akt and the ratio of NF-κB p65 in the nucleus versus cytoplasm,there were significant differences among LY294002 group,FSH + LY294002 group and FSH group (P ＜ 0.05 ).ConclusionThe effects of FSH on proliferation and invasion of ovarian cancer cell lines SKOV3 and 3AO may be realized by regulating the activity of NF-κB in PI3K/Akt signal pathway.