0 ^05). Giemsa positive samples were all positive by PCR-ELISA. The negative control group had one positive by ELISA in lung tissue and BALF respectively. Conclusion PCR-ELISA shows a high sensitivity and specificity in detecting the DNA of Pneumocystis carinii, which is a secure and easy use method.

Objective To investigate the effect of lentivirus-mediated siRNA interference of USP39 on proliferation and migration of mice vascular smooth muscle cell in vitro.Methods Five siRNAs of siControl, siRNAUSP39-70, siRNAUSP39-71, siRNAUSP39-72 and siRNAUSP39-73 were designed and sythezied,mice VSMCs were infected with the lentivirus for delivering siRNAUSP39-73, and the stably transfected cells were selected by puromycin.The interference efficiency of siRNAUSP39-73 was assessed with Western blot.The effect of USP39 interference on the proliferation of VSMCs was determined by cells counting and MTT assay.Transwell assay was used to detect the migration of VSMCs.Results Recombinant lentiviral vector siRNAUSP39-73 was successfully transfected into mice VSMCs.Comparing with siControl group and Normal group, USP39 protein level of siRNAUSP39-73 VSMCs were decreased(P<0.05), and the proliferation and migration ability were all inhibited(P<0.05).Conclusion Targeted down-regulation of USP39 expression can inhibit the proliferation and migration of mice VSMCs in vitro.

Objective To verify the application value of the Oxford classification in child IgA nephropathy (IgAN) .Methods The clinical and pathological data by renal biospy in 123 children patients with IgAN from January 2010 to September 2013 were collected and retrospectively analyzed .84 cases were followed up .The results were divided into 4 grades(A ,B ,C ,D) based on the manifestations at the end of follow‐up .Finally the pathological analysis was performed .Results Among 123 cases ,the clinical man‐ifestations were dominated by nephrotic syndrome (42 .28% ) ,followed by hematuria complicating proteinuria (24 .39% ) .The scores of 4 pathological indexes were dominated by M 1 (82 .11% ) ,E1 (53 .66% ) ,S0 (59 .35% ) and T0 (82 .11% ) respectively ;the mesangial cells proliferation and endocapillary proliferation were related with the hematuria severity (P<0 .01);mesangial cells pro‐liferation ,endocapillary proliferation and renal tubule atrophy/interstitial fibrosis were related with the edema occurrence ( P<0 .05);the mesangial cells proliferation ,segmental glomerulosclerosis and renal tubule atrophy/interstitial fibrosis were related with the average arterial pressure increase(P<0 .05) .4 pathological indexes were related with 24 h urinary protein amount(P<0 .01);the segmental glomerulosclerosis and renal tubule atrophy/interstitial fibrosis were related with the decrease of the estimated glo‐merular filtration rate(P<0 .01) .84 cases were successfully followed up ,the clinical outcome was grade A in 43 cases(51 .19% ) , grade B in 30 cases(31 .71% ) ,grade C in 8 cases(9 .52% ) and grade D in 3 cases(3 .57% ) .Only the renal tubule atrophy/intersti‐tial fibrosis was related with prognosis(P<0 .05) .Conclusion The Oxford classification has certain relation with clinical indexes of children with IgAN .Only the renal tubule atrophy/interstitial fibrosis are the risk factors of prognosis .

Objective To investigate bladder anatomical changes and dose variation in patients with cervical cancer.Methods We analyzed 20 patients,undergoing external beam radiotherapy scanning cone beam CT(CBCT)before each fraction.Bladder was contoured on each CBCT,was projected onto the planning CT and assesses anatomical changes and dose variation.Results A total 451 CBCT images,for 20 patients were collected for analysis,show more change in bladder volume and position.In 15 cases bladder volume and V45 had no significant correlation(r=0.225 -0.473,all P>0.05),4 cases shows negative correlation(r=-0.564,P<0.05;r=-0.597,P<0.01;r=-0.942,P<0.01;r=-0.816,P<0.01),1 case shows positive correlation(r=0.662,P<0.01).Have more than the criteria(V45≤50%)number is 64/451(14.2%)in whole treatment.Conclusions For most patients by filling adequacy bladder,bladder dose variation is acceptable:CTV lager for individual patients should be closely observed its regression,implementation of the offline or online calibration.

ObjectiveTo investigate the anatomical changes and dose variation of rectum during radiotherapy in patients with cervical cancer.Methods Ten patients with cervical cancer underwent intensity-modulated radiotherapy using online cone beam computed tomography (CBCT) before each fraction.Rectum was contoured on each CBCT and projected onto the planning CT to analyze the changes of the rectal volume and position.The rectal volume receiving ≥ 45 Gy ( V45 ) was evaluated accordingly.Results227 CBCT images in 10 patients were collected.The rectal volume changed from ( 35.0 ± 7.3)cm3 to (97.7±14.7) cm3.The shift of rectal center was (0.14 ±0.06) cm in left and right direction,(0.24±0.10) cm in anterior and posterior direction,and (0.55±0.28) cm in superior and inferior direction.The V45 of rectum varied from (9.19±2.46)％ to (60.54 ±11.67)％.In7 of the 10 patients,rectal volume and V45 of the rectum had significant positive correlation (r =0.582 - 0.743,all P ＜ 0.01 ).Among the 227 images,the V45 of rectum was ≤50％ in 68 images (30.0％ ).ConclusionsSignificant changes in rectal volume and position occurred during fractionated radiotherapy in patients with cervical cancer,which resuhs in variations in the dose rectum received.For most patients,rectal volume and the V45of rectum had significant positive correlation.

OBJECTIVETo provide ideal animal models for exploring pathogenesis and experimental therapy of primary malignant melanoma of the small intestine.

METHODSThe histologically intact primary and liver metastatic fragments derived from surgical specimens of one patient with metastatic malignant melanoma of the small intestine were orthotopically implanted in the small intestinal mucous layer of nude mice. The take rate, invasion and liver metastasis were observed. Morphology (light microscopy, electron microscopy), immunophenotype analysis, flow cytometry and karyotype analysis were applied for the original human tumors and the transplanted tumors.

RESULTSThe primary and liver metastatic fragments of malignant melanoma of the small intestine were successfully implanted in nude mice. After continuous passages in nude mice,an orthotopic model of human primary malignant melanoma of the small intestine(from the primary focus)in nude mice (termed HSIM-0501) and a liver metastatic model of human primary malignant melanoma of the small intestine (from the liver metastatic focus) in nude mice (termed HSIM-0502) were established. Histological examination of transplanted tumors revealed high-grade melanoma. S-100 protein and HMB45 were positive. Massive melanin granules and melanin complex were seen in cytoplasm of tumor cells.Chromosomal modal number was between 55 and 59. DNA index (DI) was 1.49-1.61, representing heteroploid. HSIM-0501 and HSIM-0502 were maintained for 25 and 27 passages in nude mice respectively. Three hundred and seventeen nude mice were used for transplantation. Both the take rate after transplantation and resuscitation rate of liquid nitrogen cryopreservation were 100%. HSIM-0501 exhibited 46.2% liver metastasis and 36.7% lymph node metastases. In HSIM-0502, both liver and lymph node metastases were 100%.The transplanted tumors autonomically and invasively grew in the small intestines of nude mice and hematogenous metastasis, lymph node metastasis and celiac planting metastasis occurred.

CONCLUSIONTwo nude mice liver metastatic models of human primary malignant melanoma of the small intestine are successfully established, which provide ideal animal models for the research of pathogenesis,metastasis biology and anti-metastatic experimental therapy of primary malignant melanoma of the small intestine.

Animals ; Female ; Humans ; Intestinal Neoplasms ; pathology ; Intestine, Small ; Liver Neoplasms, Experimental ; secondary ; Male ; Melanoma ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation

ObjectiveTo study the chemical constituents from the roots of Flemingia philippinensis.MethodsThe chemical constituents were isolated and purified by combination of silica gel column,Sephadex LH-20,polyamide,and ODS column chromatography.The structures of the isolated compounds were identified by means of spectral data.ResultsTen compounds were isolated from F.philippinensis and identified as isoderrone (1),dalparvin A (2),prunetin (3),7,3'-dihydroxy-5,4',5'-trimethoxyisoflavone (4),pratensein-7-O-β-D-glucoside (5),sissotrin (6),sophororicoside (7),formononetin (8),orobol (9),and biochanin A (10).ConclusionCompounds 1-6 are obtained from this plant for the first time.

Objective To study the relationship between status of methylation of tumor suppressor candidate 1 gene(TUSC1) promoter and expression of its protein in adolescent papillary thyroid carcinoma(PTC). Methods Forty cases of adolescent PTC were chosen and the corresponding para carcinoma tissues were taken from July 2010 to Decem-ber 2013 in the First Affiliated Hospital of Zhengzhou University surgical specimens of the thyroid gland and were con-firmed by pathology. Male 12 cases,female 28 cases,median age 14 (10-18) years old. Tumor node metastasis (TNM) stageⅠ-Ⅱ13 cases,Ⅲ-Ⅳstage 27 cases;gradeⅠin 15 cases,gradeⅡin 25 cases;lymph node metastasis in 22 cases,18 cases were negative. Methylation-specific polymerase chain reaction (MSP) and Western blot were applied respectively to examine the methyaltion of TUSC1 gene promoter and its protein expression of 40 samples of adolescent PTC and their matched adjacent non-cancerous epithelium. Results The results of MSP revealed that there was no methylation of TUSC1 gene promoter in adjacent non-cancerous epithelium,while in the adolescent PTC,the hyper-methylation rate was 60%(24/60 cases,χ2=34. 28,P<0. 05). In additional,it was related to the TNM stage,pathological grade and lymph node metastasis (χ2=4. 862,7. 111,5. 625,all P<0. 05). The result of Western blot revealed that the positive expression rate of TUSC1 protein was 100% in adjacent non-cancerous epithelium and 30%(12/40 cases) in adolescent PTC (χ2=14. 118,P<0. 05),which was related to the TNM stage,pathological grade and lymph node metastasis (χ2=5. 215,6. 222, 5. 079,all P<0. 05). There was distinct correlation between methylation of TUSC1 gene promoter and the protein expres-sion (r=-0. 84,P<0. 05). Conclusions Methylation of promoter might be one of the important mechanisms of inactiva-tion of TUSC1 gene,and might play an important role in carcinogenesis and progression of adolescent PTC.

Objective To explore the effect of diammonium glycyrrhizinate(DG) and astragalus membranaceus (AM) injection on the clinical comprehensive score in patients with acute lung injury (ALI). Methods According to the random number table method,a prospective random controlled study was conducted in which 60 cases of patients with ALI were divided into a study group and a control group(each,30 cases). Both groups received a comprehensive treatment based on the new guidelines,and the study group was additionally given DG and AM injection(DG 150 mg+AM 20 ml)one time per day for 7 days. The scores of lung injury,acute physiology and chronic health evaluationⅡ(APACHEⅡ)and systemic inflammatory response syndrome(SIRS)were measured at baseline,3rd and 7th day after treatment,and ventilation support time and final disease mortality rate were also calculated in all the patients. Results There were no statistically significant differences between the two groups in the scores of lung injury,APACHEⅡand SIRS before treatment and after treatment for 3 days(all P>0.05),with prolonged treatment,the above indexes were significantly reduced compared with those before treatment in the two groups,and the decreases in scores of indexes in study group was more significant than those in control group after treatment(lung injury score:1.31±0.99 vs. 2.29±1.08,APACHEⅡscore:18.43±8.17 vs. 24.23±6.98,SIRS score:1.69±0.89 vs. 2.60±1.04,all P<0.01). The time(hour)for ventilator support in study group was shorter than that in the control group(176.10±57.81 vs. 286.07 ± 156.27,P<0.01),but there was no statistically significant difference in mortality rate between the two groups(13.33%vs.16.67%,P>0.05). Conclusion The results suggest that DG and AM injection improve the scores of lung injury,APACHEⅡand SIRS,and alleviate the lung injury,so that the injection is beneficial to the early weaning from the ventilator to support treatment in patients with acute lung injury,and has certain therapeutic effect on ALI.

Objective To make staining preparation of tapeworm for the purpose of teaching and scientific research.Methoeds Pregnant segmens were stained by Indian ink,and mature segments,cephalomeres and bladder worms were stained by carmine.Result After staining the characteristics of pregnant segments,mature segments,cephalomeres and bladder worms were observed obviously.For instance,the stem and branches linking both-side of uteri in pregnant segment and reproduction cavity projecting in one of edges in the center of the pregnant segment were clearly shown in ink colour after stained by Indian ink.The red colour was shown in the mature segment with deep-coloured male and female reproduction systems inside,separately,after stained by carmine.Conclusion Tapeworm preparation stained by Indian ink and carmine is useful and can be preserved permanently.