Objective To study the effects of midkine (MK) gene-targeting small interfering RNA (siRNA)on the invasion of melanoma cells.Methods Three MK gene-targeting siRNAs (S1,S2 and S3)were designed,constructed,and transfected into human A375 melanoma cells.Real-time PCR was performed to measure the expression of MK gene and to screen the siRNA with best efficacy.Then,A375 cells were transfected with the optimal siRNA of various doses (3.125,6.25 and 12.5 nmol/L)followed by additional culture of various durations(24,48,72 hours).Some A375 cells remaining untreated served as the blank control group,and some transfected only with liposomes served as the vector control group.Reverse transcription (RT) -PCR and Western blot were conducted to detect the mRNA and protein expression of MK,respectively,MTT assay to observe the adhesion of A375 cells,and Boyden chamber was used to evaluate cell invasion.Results The expression of MK mRNA was downregulated by all the three siRNAs,especially by the siRNA S3,which was used in the following transfection experiment.Real-time quantitative PCR revealed that the MK mRNA expression was reduced by the siRNA in a dose- (r24hours=-0.906,r4Bhours=-0.922,r72hours=-0.939,all P<0.01)and time-dependent(r3.125nmol/L=-0.889,r625nmol/L=-0.935,r125nmol/L=-0.928,all P<0.01)manner.MTT assay showed that the percentage of adhesing cells was 73.66%±2.25%,49.36%±2.16%and 28.35%±1.68%in A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively.The number of cells migrating across the chamber filter was 23.9±1.6,12.1±1.5,5.6±1.2 among A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively,significantly lower than that in the blank control group(36.8±1.5).The percentage of adhesing cells and number of migrating cells decreased with the dose of siRNA(r=-0.936,-0.915,P<0.01,0.05,respectively).Conclusions MK gene might play an important role in the adhesion and invasion of melanonla cells.To down-regulate the expression of MK gene by siRNA may suppress the adhesion and invasion of melanoma cells.

Objective To explore the clinical characteristics,misdiagnosis reason,diagnosis and treatment of primany small intestinal tumor.Methoed Retrospective analysis of the clinical data were made on 40 cases of primary small intestinal tumors.Results Before operation,13 cases were given confirmed diagnosis of small intestinal tumor.The rate of misdiagnosis was 67.5％ (27/40).Partial excision were administered in 10 cases with benign small intestinal tumor.Adical excaision were performed in 24 cases with malignant tumors,palliative excision in 3 cases and shortcut operation in 3 cases.One patient died after opreation.Conclusions Before operation,the misdiagnostic rate of primary small intestinal tumor is high.Fiber endoscopy and X-ray may bring a high diagnostic rate of small intestinal tumor.

Objective To explore the correlation of mycoplasma pneumoniae infection with frequent relapses of steroid-sensitive nephrotic syndrome (SSNS) in children.Methods 35 patients with relapse of SSNS and acute respiratory tract infection were divided into a observation group (mycoplasma pneumoniae infection) and a control group.The clinical and laboratory data including 24 h urine protein (24 h-Upro),urea nitrogen (Bun),serum creatinine (Scr),albumin (Alb) and cholesterol (Chol) were analyzed before and after treatment.Results The clinical and laboratory indexes were obviously improved after treatment,the difference was statistically significant (P ≤ 0.01).24 h-Upro decreased more significantly in the observation group than in the control group after treatment.In the observation group,15 of 18 children achieved the efficacy,9 of whom had complete response and 6 had partial response.In the control group,14 patients achieved the efficacy,6 of whom had complete response and 8 had partial response.Conclusions After treatment,most of the children with frequent relapses of steroid-sensitive nephrotic syndrome induced by cute respiratory infection are relieved.Proteinuria,hypoproteinemia,hyperlipidemia,and renal function were improved in those patients.Therapies with azithromycin achieves a more marked efficacy.

Objective To investigate the effect of mitogen-activated protein kinase(MAPK)pathway on the transcriptional expression of mdr1 gene induced by doxorubicin ( DOX)and study the transcription regulation of mdr1 gene.Methods K562 cells were treated with DOX(0.01 μg/ml)with the initial concentration of 0.01 μg/ml for 24 hours,then change the culture media without DOX.K562 cells were cultured until the its status wag recovered.Subsequently the cells were treated with DOX(0.02μg/ml)for 24 hours again.The concentration of DOX was increaged until 0.05 μg/ml by following the protocol above.K562 cells were collected at the concentration of 0.01 μg/ml,0.03μg/ml and 0.05μS/ml DOX.Expression of mdr1 gene were examined by reverse transcription-polymerase chain reaction(RT-PCR).Pglycoprotein(P-gP)wag detected by flow cytometry.Western blot wag performed to detect ERK and P-ERk.K562 cells were pretreated with MAPK inhibitor PD98059 for 1 hour.and then DOX was added.RT-PCR and FCM were used to detect the expression of mdr1 mRNA and P-gp.Results When K562 cells were exposured to DOX.the phosphorylation of ERK wag increaged.the mdr1 gene wag highly expressed as well as its corresponding protein P-gp.When the concentration of DOX was 0.05μg/ml,the expression of mdr1 gene and P-gp were increased over 5 fold.When K562 cells were pretreated with MAPK inhibitor PD98059,the expression of mdr1 gene induced by DOX(the concentration was 0.03 μg/ml and 0.05 μg/m1)was effectively inhibited by(74.1±0.11)%and(70.2±0.14)%respectively.Conclusions DOX could induce the expression of mdr1 gene in K562 cells accompanied by the activation of MAPK/ERK pathway.The block of activation of ERK could inhibit the induced expression of mdr1 gene.

ObjectiveTo study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA(siRNA) on adhesion and invasion of human breast cancer cell. MethodsReal time PCR was used to evaluate the TROP-2 mRNA of seven human breast cancer cell lines Bcap-37 ,LCC1 ,MCF-7 ,MDA-MB-231,MDA-MB-435, MDA-MB-468 ,and ZR75-1. The cell line of TROP-2 highest expression was transfected with different dose of TROP-2 siRNA. The expression of TROP-2 mRNA and protein were determined by Real-time quantitative PCR and immumoflurescence method. The cell adhesion was evaluated by MTT assay,and invasion was exmined by hoyden chamber,respectively. Results Cell line MCF-7 showed the highest elevation of TROP-2 mRNA in seven breast cancer cell lines. The results from real-time quantitative PCR and immumoflurescence method showed that TROP-2 mRNA and protein reduced in time-and dose-dependent manners( P ＜ 0.01 ;P ＜ 0.01 ). The adhesive rate of siRNA groups(5 nM,10 nM,and 20 nM)was(52.9 +2.5)％ ,(25.6 ±2.3)％, ( 12.8 +2.2)％ (P ＜0.01 ) ,respectively.The transwell results showed that the invasion cells was(78 ± 17), (39 ± 15), ( 19 ± 16), ( 136 +25 ) and( 139 ±21 )in different groups(5,10,20 nM siRNA,and controls) ,respectively(P ＜0.01). ConclusionTROP-2 gene might play an important role in adhesion and invasion of human breast cancer cell. siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cell.

Objective To comparative study the effect on colorectal cleansing of CT colonography with gulping down 10 mg bisacodyl before or 1 h after oral taking 2 liter polyethylene glycol .Methods Forty participants with informed consent were appor‐tioned to group A ,group B randomly ,20 cases in each group .On the day before CT colonography ,participants in group A oral took 20 mL of 40% W/V barium sulfate prior to 3 mealtime ,and 20 mL of 60% diatrizoate meglumine diluted in 250 mL of water after supper ,then gulped down 10 mg bisacodyl enteric‐coated tablets 1 hour before oral taking 2 liter polyethylene glycol electrolyte so‐lution .Participants in group B were the same as that in group A ,with the exception of gulping down 10 mg bisacodyl enteric‐coated tablets 1 hour after oral taking 2 liter polyethylene glycol electrolyte solution .Cleansing efficacy of stool and fluid ,and attenuation value of remainder fluid between the two groups were analyzed statistically .Results In group A ,score of cleansing efficacy of stool (1 .96 ± 0 .11) was lower than that in group B (2 .01 ± 0 .12) ,segments with good cleansing efficacy of stool (87/120 segments , 72 .50% ) was higher than that in group B (83/120 segments ,69 .17% ) ,the difference was not statistically significant (P>0 .05) .In group A ,score of cleansing efficacy of fluid (1 .50 ± 0 .06) was lower than that in group B (1 .53 ± 0 .06) ,segments with good cleansing efficacy of fluid(113/120 segments ,94 .17% ) was higher than that in group B (111/120 segments ,92 .50% ) ,the differ‐ence was not statistically significant (P>0 .05) .Attenuation value of remainder fluid [(729 ± 29)HU ] in group A was higher than that in group B[(653 ± 25)HU] ,the difference was statistically significant(P<0 .05) .Conclusion Gulping down 10 mg Bisacodyl before or after oral taking 2 liter polyethylene glycol has no effect on cleansing of stool and fluid ,with good cleansing efficacy .The former has better cleansing efficacy of fluid ,is beneficial to detecting polyps for CT colonography .

Objective To study the effects of midkine(MK)gene small interfering RNA(siRNA)on adhesion and invasion of human breast cancer cells.Methods Real time PCR was used to evaluate MK mRNA expression in 7 human breast cancer cell lines Bcap-37,LCCI,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,and ZR75-1.The cell line in which MK expression was the highest was transfected with different doses of MK siRNA.The expression of MK mRNA and protein was determined by real-time quantitative PCR and immunoflurescence staining.The cell adhesion was evaluated by MTT assay and invasion was examined by Boyden chamber method.Results Cell line MCF-7 expressed the highestlevel of MK mRNA in the 7 tested breast cancer cell lines.After being transfected with MK siRNA,MK mRNA and protein level of MCF-7 decreased in timeand dose-dependent manners.The adhesive and invasive ability of MCF-7 cell transfected with MK siRNA decreased in a dose dependent manner(P<0.01,P<0.01).Conclusions MK gene might play an important role in adhesion and invasion of human breast cancer cells.siRNA transfection could effectively inhibit adhesion,migration,and invasion of human breast cancer cell.

OBJECTIVETo evaluate the clinical outcome of craniofacial resection for advanced malignant tumors in oral and maxillofacial regions.

METHODSForty-six patients who underwent craniofacial resection for malignancies involving the anterior and middle cranial fossa over a 20-year period between June 1978 and December 1997 at our department were evaluated. Twenty patients received radiation therapy and an adjuvant therapy after the operation. Eleven patients received chemotherapy of various types as an adjuvant therapy.

RESULTSThe 3- and 5-year survival rates were 48.8% (20/41) and 35.1% (13/37), respectively, while the 10-year survival rate was 20% (4/20).

CONCLUSIONSOur results revealed good prospects of using craniofacial resection on patients with advanced malignancies in the oral and maxillofacial regions.

Adolescent ; Adult ; Aged ; Combined Modality Therapy ; Facial Bones ; surgery ; Female ; Humans ; Male ; Maxillary Neoplasms ; surgery ; Middle Aged ; Mouth Neoplasms ; surgery ; Skull ; surgery ; Skull Neoplasms ; surgery