Objective To investigate four methods for establishing animal models of human breast cancer bone metastasis. Methods Thirty-two female nude mice aged 4-6 weeks were divided randomly into four groups (n=8 in each group). 5×105 MDA-MB-231 cells were injected into the body via the left second mammary fat pads (group A), the tail veins (group B), the left heart ventricles (group C) and the left tibia marrow cavities (group D), respectively. Tumor formations in situ were recorded in group A. Deaths after the injection were recorded. The surviving nude mice 49 days after the injection were subjected to pathological examination to determine bone metastasis. Results The rate of tumor formation in situ of group A was87.5 %(7/8). One mouse in group C died after the injection of MDA-MB-231 cells. The bone metastasis rate in groups A, B, C and D was zero (0/8), 12.5 % (1/8), 71.4 % (5/7) and 100 % (8/8), respectively. There was statistically significant difference in the bone metastasis rate between group A and group C, group A and group D, group B and group C; and group B and group D. Conclusion Injections of tumor cells via the breast fat pads and tail veins were not suitablemethods to establish animal models of human breast cancer bone metastasis. The bone metastasis model could be established efficiently by injecting tumor cells into the left heart ventricles or the bone marrow cavity of nude mice.

Objective To apply a fluorescent nucleotide dye Sybr Green I in detecting telomerase activity and assess its clinical value in the diagnosis of colorectal carcinoma. Methods Telomerase activity was (measured) by telomeric repeat amplification protocol (TRAP) combined with fluorescent nucleotide dye Sybr Green I in 46 cases of colorectal carcinoma tissue specimens and 19 tumor-adjacent tissue (specimens). Results (Telomeric) repeat amplification product was displayed as clear bands in PAGE stained with Sybr Green I; the positive rate of telomoerase activity in colorectal carcinoma tissue was 89.13%, whereas no telomerase (activity) was observed in 19 tumor-adjacent tissue specimens. There was no (relationship) between telomerase (activity) and the level of cell differentiation, Dukes stages, or metastasis of lymph nodes. Conclusions The method of TRAP combined with Sybr Green I to determine telemerase (activity) is an important method for the diagnosis of colorectal carcinoma.

Objective To investigate the utility of serum prostate specific antigen(PSA) density (PSAD),prostate antigen transition zone density(PSAT) and the ratio of free/total PSA with PSAD [(F/T)/PSAD] in the diagnosis of prostatic carcinoma (PCa) by three-dimensional ultrasonography.Methods Seventy-eight patients (serum prostate-specific antigen between 4-20 μg/L ) were involved.The prostatic volume and its transition zone volume were measured by three-dimensional ultrasonography.Then the relative parameters of PSA [PSAD,PSAT and (F/T)/PSAD] were calculated.Pathologic types were determined by using needle biopsy of prostate.Results Among them,27 patients were suffering from PCa,while the other 51 benign prostate hypertrophy (BPH).The difference of PSAD,PSAT and (F/T)/PSAD between PCa and BPH had arrived statistical significance (P<0.05).Proportions under the PCa curves were 0.736,0.760,0.800 respectively.Considering both sensitivity and specificity,a cutoff was recommanded:PSAD>0.20,PSAT>0.33,(F/T)/PSAD<0.8.Conclusions When the serum PSA level is between 4 μg/L and 20 μg/L,PSAD,PSAT and (F/T)/PSAD are of significant value to differentiate PCa from benign prostatic hyperplasia patients.The data are more reliable if prostatic volume are calculated by three-dimensional transrectal ultrasonography.

Objective To assess the application value of REP-PCR in genotyping of candida glabrata strains in clinical pratice. MethodsFrom 2009 to 2010, thirty-eight candida glabrata strains were isolated from Shanghai Ruijin Hospitals, Shanghai Renji Hospital, Shanghai Huashan Hospital, Anhui Medical University Hospital, Shenzhen People's Hospital. Six loci in housekeeping genes (FKS, LEU2,NMT1, TRP1, UGP1 and URA3 ) were amplified and sequenced. The sequences were compared with the MIST database and allele profile and sequence type (ST) were obtained. With primers Ca21, Ca22 and Com21 used to amplify the adjacent variable gene regions,the amplicons were analyzed through electrophoresis to generate different REP-PCR types. Finally, the results of these two genotyping methods were compared. ResultsFor REP-PCR, Ca22-Com21 has the best genotyping effect. REP-PCR and MLST have the same genotyping results. Five REP-PCR types were found in 38 candida glabrsta isolates. Type A,B, C, D and E strains from REP-PCR were genotyped as ST 7, 3, 19, 45 and new type respectively byMIST. REP-PCR saves time compared with MIST. Conclusions REP-PCR offers a simple and rapid method for molecular typing, with similar discriminatory power with MIST. Therefore, REP-PCR can be the preferred choice in laboratory, especially for a large number of isolates.

Objective To determine the effect of NEL-like type 1 gene (NELL-1) transfection in vivo in the repair of traumatic femoral head necrosis.Methods Twenty-four SD rats were randomly divided into three groups (8 rats per group) according to the lottery method,ie,sham group (served as normal control),NELL-1 treatment group (injected NELL-1 gene by recombinant adenovirus vectors around the hip one week after osteonecrosis model induced surgically) and placebo group (given an equal volume of saline solution at the same time after the induction of osteonecrosis).Femurs were taken from the animals 5 weeks after surgery.Gross observation was performed for morphology changes,X-ray assessment for femoral head height and length ratio (H/L),Micro-CT measure for bone parameters of femoral head including total volume (TV),bone volume (BV),total mineralized content (TMC),trabecular thickness (Tb.Th) and trabecular space (Tb.SP),and histological study for osteocytes,osteoblasts and osteoclasts.Results Preserved femoral head shape was noted in NELL-1 treatment group compared to the obvious flattening of the femoral head in placebo group.No heterotopic osteogenesis was observed in any group.Femoral head H/L ratio for 0.753 2 ± 0.040 2 in NELL-1 treatment group was higher than 0.598 4 ± 0.037 0 in placebo group (P ＜ 0.05),but lower than 0.920 2 ± 0.037 0 in sham group (P＜0.05).TV,BV,TMC and BMD between NELL-1 treatment and sham groups did not differ significantly (P ＞ 0.05),but all were increased compared to placebo group (P ＜ 0.05).There was no significant differences in Tb.Th and Tb.SP among three groups (P ＞ 0.05).Most osteocytes were alive in NELL-1 treatment group.More active osteoblasts and osteoclasts were noted in NELL-1 treatment group than those in placebo group.Conclusion NELL-1 gene transfection can preserve femoral head shape and bone content,promote osteoblast activity and neovascularization and hence is an effective treatment for rat traumatic osteonecrosis.However,the activity of osteoclasts is stimulated simultaneously.

OBJECTIVETo observe the efficacy and safety of Qizhi Jiangtang Capsule (QJC) in treating stage 3b diabetic kidney disease (DKD) patients with macroalbuminuria.

METHODSPatients who conformed to the diagnostic criteria of stage 3b DKD were randomly assigned to two groups according to random digital table, the experiment group and the control group, 84 in each group. All patients received a two-week elution period, and then were treated with basic Western therapy. Patients in the experiment group took QJC, 5 pills per time, 3 times a day, while those in the control group took Valsartan Capsule 160 mg each time, once daily. The observation period of follow-ups was limited within 6 months, and the time points were set as the baseline, 1st month, 3rd month, and 6th month. Systolic blood pressure (SBP), diastolic blood pressure (DBS), 24 h urine protein quantitative (24 h UPQ), plasma albumin (ALB), and serum creatinine (SCr) were detected and recorded, and estimated glomerular filtration rate (eGFR) was calculated. The occurrence of hypoglycemic reaction, coagulation disorder, gastrointestinal tract reaction, allergy, hyperkalemia, doubling of creatinine, and overall adverse events were observed and recorded at same time.

RESULTSFinally 81 patients in the experiment group and 80 patients in the control group were effectively included. Compared with the baseline level, SBP and DBS obviously decreased in the control group at month 1 of treatment (P < 0.05), and more significantly decreased at month 6 of treatment (P < 0.01). SBP at month 1, 3, and 6 of follow-ups; DBS at month 6 of follow-ups was lower in the control group than in the experiment group (P < 0.05). At month 1, 3, and 6 of follow-ups, 24 h UPQ of the experiment group was significantly lower than the baseline level (P < 0.01). It was also significantly lower than the level of the control group at the same time point (P < 0.05). There was no significant difference in 24 h UPQ at month 1, 3, and 6 of follow-ups between the control group and the baseline level (P > 0.05). ALB of the experiment group showed an increasing trend. It was significantly higher than the baseline level at month 6 (P < 0.05), which was also higher than that of the control group at same period (P < 0.05). There was no significant difference in the ALB level in the control group (P > 0.05). SCr of two groups showed an increasing trend. SCr of the experiment group was significantly higher at month 1, 3, and 6 follow-ups than the baseline level (P < 0.05). But the increment of SCr was higher in the control group than in the experimental group, and obviously higher than the baseline levels (P < 0.05). eGFR of both groups showed a decreasing trend. The decrement was higher in the control group than in the experimental group (P < 0.05). The proportion of progression of renal functions at month 1, 3, and 6 of follow-ups in the experimental group was 0.0% (0 case), 9.55% (8 cases), and 21.4% (18 cases), while they were 8.3% (7 cases), 21.4% (18 cases), and 40.5% (34 cases) in the control group. There was no statistical difference in the proportion of progression of renal functions between the two groups at month 3 and 6 of follow-ups (P < 0.05). There was no statistical difference in the incidence of adverse reactions between two groups (P > 0.05).

CONCLUSIONQJC could effectively reduce urinary protein of patients with stage 3b DKD, and delay the progression of renal functions.

Adult ; Albumins ; analysis ; Albuminuria ; drug therapy ; Blood Pressure ; drug effects ; Creatinine ; blood ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerular Filtration Rate ; Humans ; Male ; Middle Aged ; Tetrazoles ; therapeutic use ; Treatment Outcome ; Valine ; analogs & derivatives ; therapeutic use ; Valsartan

Osteonecrosis is a common disease, mainly affecting femoral head. Good animal models are helpful in research on the pathologic mechanism of osteonecrosis and the exploration of effective treatment. Although it is relatively easy to establish animal models of early osteonecrosis of femoral head using various approaches, it is difficult to develop an animal model that mimics the full range of osteonecrosis of femoral head. In this paper, we reviewed the current researches on experimental animal models of osteonecrosis, with an attempt to provide evidences for choosing the appropriate animal models and find the way of future development.
Animals ; Disease Models, Animal ; Osteonecrosis

Objective To explore the value of BI-RADS scoring system based on the sonographic features in the breast nodules.Methods In order to build a Logistic regression model,regression was made to analyse 401 patients ' sonographic features of breast nodules.A scoring system was developed based on the results of regression's filter and the weight of each factor,used to score and classify the other 243 patients.It's diagnostic value was evaluated through comparing all types of theoretic risk ranges of BI-RADS.Results Age,morphology,orientation,margin,echo pattern and microcalcifications in a mass were selected in the final step of the logistic regression analysis.By means of scoring system,the scores corresponding to BI-RADS 3,4a,4b,4c,5 classes are 6,7-8,9-15,16-22 and ≥23 respectively.Case study comprehensive score of BI-RADS 3-5 classification' s positive predictive values were 0,4.17％,21.43％,84.85％,1 00％,and the area under the ROC curve scoring system was 0.947.Conclusions The scoring system can objectively score and classify breast nodules,and therefore provide an effective reference for clinical evaluation of benign and malignant breast.

Objective To investigate the effects of miR-10a expression on migration and invasion of human pancreatic cancer cells AsPC-1.Methods Small interfering RNA targeting at miR-10a (miR-10a-siRNA) was constructed,then it was transfected into pancreatic cancer AsPC-1 cells,and nonsense siRNA (Nc-siRNA) group and blank control group was established.Real time PCR assay was used to detect the expression of miR-10a in the 3 groups,and wound healing assay and Transwell assay were used to determine the migration and invasion abilities of cancer cells.The amount of matrix metalloproteinase-13 (MMP-13) in supernatant of cancer cell culture of each group was examined by ELISA assay.Results The miR-10a levels in control group,NC-siRNA group and miR-10a-siRNA group were 1.05 ±0.08,1.03 ±0.06,0.02 ±0.01 ; and the number of transmembrane cell were (150 ± 2.6),(145 ± 2.2),(62 ± 1.8),the levels of MMP-13 in the supernatant were (108.5 ± 2.8),(107.8 ± 2.5),(35.8 ± 1.5) pg/ml.The values were significantly lower in miR-10a-siRNA group than those in control group and NC-siRNA group (P ＜ 0.01).The distance of cultured clone in miR-10a treated cancer cells (736± 18 μm) was significantly longer than those in the controls (385 ±5 μm) and NC-siRNA group (395± 13 μm,P＜0.01).Conclusions Down-regulation of miR-10a by siRNA may inhibit migration and invasion of pancreatic cancer AsPC-1 cells,and the downregulated expression of MMP-13 may be one of the important mechanisms.

ObjectiveTo evaluate the application of chest-abdomen longitudinal diameter ratio,total lung area and lung longitudinal diameter in different gestational ages by two-dimensional ultrasonography.MethodsFive hundred and ninty-two fetus of normal singleton pregnancy between 18 and 40 weeks were ultrasonic scanned by standard 4-chamber view and fetus torso coronal section.The heart area,thoracic area,bilateral lungs pointed to the corresponding diaphragmatic dome gallery on the top of the vertical distance,diaphragmatic dome iterated top to the bottom of the fetal bladder distance were measured respectively.Related regression analysis was done on fitting total lung area and longitudinal diameter with pregnant increasing.ResultsA normal pregnancy fetal total lung area and lung longitudinal diameter were increased as long as the gestational age.The best-fit regression equations of total lung area was:Y =0.83 X +13.894,R2 =0.914;Lung longitudinal diameter was:Y =0.669 X + 3.124,R2 =0.892.Chest-abdomen longitudinal diameter ratio from 18 to 40 weeks gestation was in constant range:0.44 - 0.59.Conclusions Chest-abdomen longitudinal diameter ratio from 18 to 40 weeks gestation is in constant range.The normal pregnancy fetal total lung area and lung longitudinal diameter were increased as long as the gestational age.These parameters may be useful for the prenatal assessment of lung development.