Objective To observe the effect of isolating and harvesting the chondrocytes from rabbits rib cartilage with the method of three-step enzymatic digestion, and the biological characteristic of the isolated chondrocytes during cultivation in vitro to evaluate their biological activity. Methods The method of three-step enzymatic digestion was designed that the rib cartilage was digested one by one with 1 g/L trypsin and 1 g/L EDTA, 1 g/L hyaluronidase and 2 g/L collagenaseⅠ in the culture medium to isolate chondrocytes. The harvesting and viability rate of the primary chondrocytes were detected. During the passage cultivation in vitro, the changes of the chondrocyte shape and growth were observed, and the changes of the collagen typeⅠ and Ⅱ and aggrecan in the extracellular matrix were detected. Results ① The extracellular matrix of rib cartilage was completely dissolved by the three-step enzymatic digestion, and the chondrocytes were completely isolated from the solid matrix. The number of the harvested chondrocytes from every gram of wet cartilage was (4 295.7)?10~(4) on average,and their viability rate was 97.2% on average. ②The primary and first passage chondrocytes had triangle or multi-angle shape, and became elliptic shape at the growing confluence with the positive immunohistochemical staining of collagen type Ⅱ and the strong heterochromia to toluidine blue. The content of sulfate glycosaminoglycans(GAG) in the extracellular matrix of the primary passage cells was (80.61?11.40) ?g/cm~(2). The chondrocytes after the third passage gradually became spindle shape with the negative staining of collagen typeⅡ and the weak heterochromia to toluidine blue. The content of sulfate GAG of the fourth passage cells was (44.74?10.18) ?g/cm~(2). Conclusion ① The method of three-step enzymatic digestion can make the extracellular matix of rib cartilage to be completely degraded, and has advantages of the high efficiency of harvesting primary chondrocytes with high cellular viability rate and simple manipulation. ②The primary and first passage chondrocytes have fine biological activity.

Objective To study the protein expression of Tspan-1 gene in colorectal carcinoma,and investigate the relationship between the expression and biologic behavior of colorectal carcinoma. Methods Immunohistochemical ELIVISION two-step method was used to detect the protein expression of Tspan-1 gene in 80 cases of colorectal carcinoma,13 cases of colorectal adenoma and 27 cases of normal colorectal tissues. Results The positive expression rate of Tspan-1 in colorectal carcinoma was significantly higher than that in colorectal adenoma and normal colorectal tissues,there was significant difference in the expression among the three groups.The positive expression rate of Tspan-1 was 90% in colorectal carcinoma,23% in colorectal adenoma and 7% in normal colorectal tissues (P ＜ 0.01).The positive expression rate of Tspan-1 in poor and moderately differentiated carcinoma was significantly higher than that in well-differentiated carcinoma (P ＜ 0.01).The expression of Tspan-1 was significantly associated with lymph node metastasis (P＜0.01),and with the 5-year survival rate of colorectal carcinoma patients,with also TNM stage and the depth of tumor invasion (P ＜0.01),and while it was not associated with gender,age,location,tumor size,and type (P ＞ 0.05). Conclusion The expression of Tspan-1 gene could be as a marker predicting the prodnosis of colorectal adenocarcinoma patients,also it may play an important role in the pathogenesis of colorectal adenoma.

Objective To explore risk factors relating to occurrence of spontaneous rupture (SR)of hepatocellular carcinoma (HCC). Methods We retrospectively reviewed medical records of 409 HCC patients treated in our department from Jan. 1st 2005 to Aug. 31st 2009. Clinicopathologic factors were explored for their possible association with occurrence of SR by univariate and multivariate analysis using Logistic regression model. Results Among 409 patients, SR occurred in 40 patients (9. 8% , 40/409).Type of SR was defined according to presentation of acute abdominal emergency, especially unstable hemodynamic status. Nineteen cases (4. 7%, 19/409) were grouped as type of HCC-SR Ⅰ and 21 cases (5. 1% ,21/409) as HCC-SR Ⅱ. Patients in the two groups possessed similar hepatitis infection status,severity of background liver disease and tumoral factors except for tumoral location, while HCC-SR Ⅰ group often needed emergency treatment demonstrating elevated values of aspartate aminotransferase (AST) and WBC, decreased PLT counts and worse Child-Pugh classification. Patients in the HCC-SR Ⅱ group had more opportunities of receiving curative hepatectomy and had longer overall survival. The 1,2 and 3 year survival rates were 26. 3 % (5/19), 5. 3% ( 1/19 ) and 0% (0/19)in the HCC-SR Ⅰ group and 66. 7% ( 14/21 ),42.9% (9/21)and 19. 5% (5/21) in the HCC-SR Ⅱ group, respectively. Differences reached statistical significance( P = 0. 011,0. 009,0. 049). Multivariate analysis identified that severity of concomitant liver cirrhosis , WBC ＞ 10 × 109/L and AST level more than twice normal (2N) as independent risk factors associating with occurrence of HCC-SR Ⅰ , while elevated values of AFP and presence of tumor thrombi in portal vein branch related to HCC-SR Ⅱ. Conclusions HCC-SR includes two different types, since different risk factors predict occurrence of each HCC-SR type.

Objective To analyze C4d deposition in the patients with late acute renal allograft rejection,and explore the role of C4d in grafts survival and grafts loss. Methods Thirty-six patients clinical and pathologically diagnosed as having acute rejection more than one year post-transplant were selected. C4d was detected by immunohistochemistry in renal allograft biopsies. The effect of C4d deposition on long-term graft survival was studied. Results Among 36 recipients with late acute renal allograft rejection, 16 cases were positive for C4d (44.4 %) and 20 negative for C4d (55.6 %). Five cases experienced graft loss in C4d positive group (31.3 %), while 6 cases in C4d negative group (30.0%). There was no significant difference in the graft loss rate between C4d-positive group and C4d-negative group. Log-Rank test demonstrated there was no significant difference in graft survival between C4d-positive group and C4d-negative group. The count of the interstitial infiltrated eosinophils in renal allograft was (9.4 + 4.5) and (2.6 + 1.8) respectively in the C4d-positive group and C4dnegative group (P＜0.05). Conclusion C4d deposition in peritubular capillary of the recipients with late acute renal allograft rejection might not be a prognostic marker for graft outcome.

Case-based teaching (CBT) on the basis of treatment guidelines was carried out to develop standard medical behavior, improve learning efficiency and students' ability of solving clini-cal problems. In clinical internship teaching, guidelines for common and frequently-occurring respira-tory diseases and CBT were organically combined. Cases were carefully selected and questions were meticulously set. Guidelines were thoroughly analyzed and teaching was reasonably evaluated. There-fore, students' learning interests were initiated and learning efficiency was improved. It not only trained evidence-based medicine thinking of intern students, but also improved the teaching ability of clinical teachers.

Objective To compare MR,CT,and radiography in the detection of arthropathies in patients with hemophilia.Methods Forty-one symptomic joint images in the 14 patients with hemophilia, aged from 11 to 24 years,were used in this study.Each joint had the examinations of radiography,CT and MR within one day.The severity of each joint was staged using conventional radiographic classification. Severe HA patients with stage 5 were excluded from the study.Imaging findings of soft tissue swelling, osteoporosis,epiphyseal overgrowth,joint erosion,cyst,joint space narrowing,bone marrow,joint effusion, hemorrhage,synovial hypertrophy,widened intercondylar notch as well as anterior and posterior crueiate ligaments(only for knee joint)were used for the all imaging comparison.Results The 41 symptomatic joints in 14 patients with hemophilia were classified by radiographic criteria into stage 0(n=5),stage 1(n=7),stage 2(n=6),stage 3(n=8)and stage 4(n=15).Soft tissue swelling or joint effusion was observed in 33 joints by radiographs,in 34 joints by both CT and MR.Joint erosions were demonstrated in 34 joints by MR,in 33 joints by CT and 20 joints by radiographs.Joint cysts were shown in 21 joints by MR,in 18 joints by CT and 9 joints by radiographs.Significant differences in detection of erosion and cyst were found between radiography with either CT(P0.05).MR showed improvement for detecting nlore loci of both erosion and cyst than CT and radiography,and also CT showed the improvement than radiography.Bone marrow edema 14 joints, hemon'hage in 34 joints and synovial hypertrophy in 27 joints were revealed on MR images.Conclusion MRI is superior to CT and conventional radiography in detecting the abnormal changes and should be considered as the first choice among the imaging modafities in evaluating hemophilic arthropathies.

Measurements of the magnetic properties and total contents of Cu, Cd, Pb and Fe in 30 automobile emission particulate samples indicated the presence of magnetic particles in them. The values of frequency dependent susceptibility (chi(fd)) showed the absence of superparamagnetic (SP) grains in the samples. The IRM(20 mT) (isothermal remanent magnetization at 20 mT) being linearly proportional to SIRM (saturation isothermal remanent magnetization) (R(2)=0.901), suggested that ferrimagnetic minerals were responsible for the magnetic properties of automobile emission particulates. The average contents of Cu, Cd, Pb and Fe in automobile emission particulates were 95.83, 22.14, 30.58 and 34727.31 mg/kg, respectively. Significant positive correlations exist between the magnetic parameters and the contents of Pb, Cu and Fe. The magnetic parameters of automobile emission particulates reflecting concentration of magnetic particles increased linearly with increase of Pb and Cu content, showed that the magnetic measurement could be used as a preliminary index for detection of Pb and Cu pollution.
Algorithms ; Environmental Monitoring ; methods ; Feasibility Studies ; Magnetics ; Metals, Heavy ; analysis ; chemistry ; Statistics as Topic ; Vehicle Emissions ; analysis

Objective Pathogens from the nosocomial infection have been analyzed by MALDI‐TOF microbial identification system ,to evaluate mass spectrometry analysis advantage and explore the mass spectrometry method .Methods The pathogens have been analyzed by MALDI‐TOF microbial identification system ,by compared with the VITEK‐2 compact detection in the tes‐ting time ,detection rate and the amounts of identified strains .The homology differences have been analyzed by comparison calcula‐tion of common peaks from the fingerprint spectrums .Results Thirty‐one Escherichia coli strains ,28 Klebsiella pneumonia strains and 9 unusual pathogen strains have been identified by MALDI‐TOF MS for only 1 hours .It has more advantages than VITEK‐2 in the testing time and other aspects .Conclusion Nosocomial infection of pathogen shows a point source propagation mode centering on the department .MALDI‐TOF mass spectrometry is able to rapidly and correctly identify the pathogen .MALDI‐TOF microbial i‐dentification system is expected to be the major detecting technique in the field of the pathogen monitor and resistance monitoring a ‐nalysis .

Objective To analyze the risk factors for postoperative stricture after endoscopic submucosal dissection (ESD) for early stage esophageal cancer.Methods The data of 362 patients with early esophageal cancer treated by ESD from January 2007 to February 2012 were reviewed to investigate the risk factors of postoperative stricture.Results Esophageal stricture after ESD occurred in 42 patients (11.6％)with a mean time from ESD to stricture of (58.5 ± 12.3) days.The rates of mild,median and severe stricture were 16.7％ (7/42),38.1％ (16/42) and 45.2％ (19/42),respectively.Multivariate analysis revealed that lesion range ＞ 3/4 esophageal circumference (odds ration [OR]:44.2 ; 95％ confidence interval [CI]:4.4-443.6) and tumor invasion beyond m2 (OR:14.2; 95 ％ CI:2.7-74.2) were independent risk factors.Stricture level was related to lesion's circumferential extension (relational coefficient (φ) =0.47,P ＜ 0.05) and tumor invasion depth (relational coefficient (φ) =0.647,P ＜ 0.05).Conclusion Circumferential extension and invasion depth of early esophageal cancer were independent risk factors for post-ESD esophageal stricture and related with the degree of stricture.

Objective Basing on overall survival(OS)of HBsAg-positive hepatocellular carcinoma(HCC)patients we evaluate several currently used HCC staging systems for OS predictivity.Methods This retrospective study included 775 patients with HBsAg-positive HCC treated in our department during 11-year period from Jan.2000 to Feb.2010.Clinicopathologic factors were evaluated for their possible association with OS in univariate and multivariate analysis using Cox proportional hazard model.Receiver operating characteristics(ROC)analysis with calculation of the area under the curve (AUC),sensitivity,and specificity was applied to define cutoff point values where appropriated and to assess HCC staging systems for their predictive ability of OS.Results The 1-,2-,3-and 5-year OS rates were 21.3％(165/775),9.4％(73/775),4.9％(38/775)and 1.7％(13/775),respectively.Multivariate analysis identified that severity of concomitant liver cirrhosis(B =4.519),treatment modality (B =4.888),ALT≥2N(B =4.068),portal vein tumor thrombi(B =0.537),spontaneous rupture(B =5.033)and inferior vena cava tumor thrombi(B =7.049)as independent risk factors influencing OS.NSMCS(North Sichuan Medical College Score)exhibited best performance predicting OS with AUC 0.801 (95％ CI 0.761-0.840),sensitivity of 78.8％,specificity of 69.3％ at NSMCS ≥-2.Median survival time reached statistically significant difference(13.6 mons,3.4 mons vs.1.3 mons,x2 =467.636,P =0.000).Conclusions Multiple factors determine OS in patients with HBsAg-positive HCC.NSMCS staging system demonstrates better predictability for the survival of HBsAg positive HCC patients.