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 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 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

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 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.

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 analyze the relationship between Arg -1 expression and the clinical pathologi-cal factors ,proliferation and prognosis value in patients with colorectal cancer .Methods The expression of Arg-1 was observed in normal tissues ,chronic inflammatory tissues ,and adenomas inflammatory carcinoma tissues of mice.At the same time,Arg-1 expression was observed in human colorectal cancer adjacent tissues ,inflamed tis-sues and colorectal cancer tissues .Arg-1 expressed in 20 cases colorectal inflammation -cancer model in mice . Arg-1 expressed in 20 normal colorectal tissues .Fiftheen colitis tissues and 110 colorectal cancer tissues were examined by Immunohistochemistry .Statistical analysis was used to analyze the changes of Arg -1 expression in different groups of mice and human colon tissue cases .Results Arg-1 protein expression in normal tissues of mice was gradually increased in colon ,chronic inflammatory tissues,adenomas,inflammatory carcinoma,with sta-tistical significances(P<0.05).Arg-1 expression in para -carcinoma tissue,colitis tissues,colorectal cancer tissues was gradually increased with statistical significance (P<0.05).Conclusion Arg-1 protein is associat-ed with colorectal cancer TNM stage .Arg-1 protein may be involved in occurrence and development process of inflammation-associated colon tumor and may be a candidate of proliferated and prognostic biomarker in patients with colorectal cancer .

[ ABSTRACT] AIM:To investigate the effects of maternal limb ischemic preconditioning ( LIP) on the mitochon-drial structures and functions of the hippocampal neurons induced by reoxygenation in the intrauterine distress fetal rats. METHODS:Pregnant rats (n=40) were randomly divided into 4 groups: sham (S) group, LIP group, fetal distress ( FD) group and LIP+FD group.Intrauterine ischemia model was established through the experimental design.The ultra-structure of the mitochondria in CA1 area of the hippocampus was observed .The mitochondrial membrane potential and re-active oxygen species ( ROS) were measured .The content of ATP and MDA in the hippocampus tissue was detected.The activity of Mn-SOD was observed.RESULTS:Compared with sham group, the ultrastructure of mitochondria in CA1 area of the hippocampus was damaged in FD group and LIP+FD group.The mitochondrial membrane potential, the content of ATP and the activity of Mn-SOD were decreased.However, the content of ROS and MDA was increased.Compared with FD group, the ultrastructure of mitochondria in CA1 area of the hippocampus was intact in LIP+FD group.Furthermore, the reduced mitochondrial membrane potential and ATP content were inhibited.The activity of Mn-SOD was increased, but the content of ROS and MDA was decreased in LIP+FD group.CONCLUSION:Limb ischemia preconditioning inhibits the damage the mitochondria of fetal hippocampal neurons induced by reoxygenation in the intrauterine distress fetal rats.

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).