Antigens, CD1 ; metabolism ; Child, Preschool ; Cholangitis, Sclerosing ; diagnosis ; etiology ; metabolism ; Diagnosis, Differential ; Female ; Histiocytosis, Langerhans-Cell ; complications ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Liver ; metabolism ; pathology ; S100 Proteins ; metabolism

BACKGROUND: Using low-intensity pulsed ultrasound (LIPU) to promote the repair of articular cartilage injury is very common,and we also have more options to choose the cytoskeleton, but the application conditions of LIPU and the appropriate cytoskeleton have not reached any consensus yet.OBJECTIVE: To investigate the feasibility of establishing tissue-engineered cartilage by cell-free allograft demineralized bone matrix (CFDBM) co-cultured with rabbit cartilage cells and bone marrow mesenchymal stem cells (BMSCs) in vitro, and to investigate the effect of LIPU on the cells in CFDBM.DESIGN, TIME AND SETTING: Multiple sample observation was performed at the Institute of Biomedical Engineering, Second Military Medical University of Chinese PLAfrom May to August 2009.MATERIALS: The CFDBM was prepared as modified Urist's method; the cartilage cells were obtained using mechanical disintegration and enzyme digestion; the BMSCs were separated using whole bone marrow rinsing method, purified, and amplified layer by layer.METHODS: As CFDBM With a composite of different cellular components, and whether applying LIPU stimulation, the samples were divided into four groups: chondrocyte group, BMSCs group, compound group (CFDBM was compounded with chondrocytes,BMSCs, and chondrocytes/BMSCs, respectively, without LIPU stimulation), and LIPU group (CFDBM was compounded with chondrocytes/BMSCs, and then the samples were stimulated with LIPU on the second day, 1.0 MHz frequency, 10 mW/cm~2 transient spatial intension, 20 min/d).MAIN OUTCOME MEASURES: ① the 2~(nd)-generation of cartilage cells and BMSCs were examined by immunohistochemical method; ② The CFDBM prepared as modified Urist's method was examined as HE staining; ③ The samples of four groups were examined by collagen II immunohistochemical staining on the 21~(st) day.RESULTS: ① The collagen II immunohistochemical staining of the second generation of the articular cartilage cells showed that the morphostructure was polygon, star or round, and pseudopodia extended, and the cells were rich in cytoplasm; the cytoplasm was brownish yellow, and the cell nuclear was round. ② The result of immunohistochemical staining of BMSCs showed that,CD34 was negative, CD44 and CD105 were positive. ③ In the center of CFDBM prepared as modified Urist's method, there was no obvious cell-like structure and the gap size was uniform. ④ On the 21~(st) day after combining CFDBM with cells, collagen II immunohistochemical staining demonstrated that BMSCs group was negative, chondrocyte group was weak positive, compoundgroup was positive, and the LIPU group was strongly positive.CONCLUSION: ① Biological property of the 1~(st)-3~(rd)-passage chondrocytes and BMSCs was similar to primary-cultured cells. ②Both chondrocytes and BMSCs had a highly proliferative ability in CFDBM. ③ 10 mW/cm~2 LIPU could not affect activity of BMSCs but could promote differentiation Into articular cartilage cells, and it also could not promote celt proliferation.

Objective: To study the propagation and phenotypes changes of killer cell (CD3AK cell activated by CD3 mAb in vitro. Methods: Lymph nodes taken from lung cancer patient is dissociated into single cell suspension by mechanical method and cultured in culture medium added CD3 mAb and a little dose IL-2. We analyze cell immunophenotype by flow cytometry and proliferation by trypan blue exclusion test per 2 days. Results: Immunophenotypic analysis showed that CD3AK expressing CD3, CD8, CD56, CD25 increased, and reached a peak value which is 2.33 times than before culturing in the 8 th day. Conclusion: CD3 mAb added to the culture medium can obviously activate CD3AK cell and stimulate proliferation and keep its killer activity.

OBJECTIVE:To establish a method for simultaneous determination of 5 effective components in Luohua zizhu dry extract. METHODS:UPLC-MS/MS was conducted. The separation was performed on an Zorbax Eclipse Plus C18 column with mo-bile phase of acetonitrile-water(gradient elution)at the flow rate of 0.3 mL/min. The column temperature was set at 40℃and sam-ple size was 2 μL. The analytes were detected in the multiple reaction monitoring(MRM)mode. Nitrogen was used as drying gas and atomized gas. The temperature and flow rate of drying gas were 325 ℃ and 6 L/min. The pressure of atomized gas was 45 psi. The temperature and flow rate of sheath gas were 350℃and 12 L/min. The voltage of capillary were 4000 V(+)and 3500 V(-). The voltage of nozzle was 500 V. RESULTS:The linear ranges of luteoloside,acteoside,quercetin,luteolin and rutin were 0.5048-252.4 ng/mL(r=0.9999),0.7124-356.2 ng/mL(r=0.9990),0.5094-254.7 ng/mL(r=0.9962),0.3030-151.5 ng/mL(r=0.9998) and 0.6022-301.1 ng/mL(r=0.9996),respectively. RSDs of precision,stability and reproducibility tests were all less than 3.0%. The limit of quantitation were 0.42,0.87,0.33,0.12,0.76 ng/mL. The recoveries were 97.99%-101.20%(RSD=1.3%,n=6), 96.50%-101.20%(RSD=1.7%,n=6), 94.81%-99.34%(RSD=1.7%,n=6), 97.54%-100.51%(RSD=1.2%,n=6), 93.37%-98.70%(RSD=1.9%,n=6),respectively. CONCLUSIONS:The method is simple,precise,stable and reproducible, and can be used for simultaneous determination of 5 effective components in Luohua zizhu dry extract.

Objective:To analyze the accuracy of the digital imaging fiber optic transillumination (DIFOTI) on diagnosis of caries lesions depth using DIAGNOcam system.Methods:This experiment adopted self-matching design.Seventy-four extracted teeth (molar:sixty-six,premolar:eight) with one caries lesions in proximity which were not damaged in surface marginal ridge were selected.Dental calculus and dental stains were removed from the extracted teeth for standby application.A sign was marked in the middle of the occlusal surface edge at the side of decay.Then the teeth were fixed in the standard model of dentition and cavities were adjacent with the sound tooth surface.Sticky wax was applied to seal the level of 2 mm beyond cemento-enamel junction (CEJ) in the direction of occlusion and interproximal space to imitate gingival margin and gingival papilla.The standard models of dentition was seated in imitation head mold.The lesions depth degree was looked into and checked with DIAGNOcam system.Besides,the pictures on the occlusal surfaces were recorded and saved.The sign above could be seen on the picture.The measuring tool in DIAGNOcam system was used to measure the depth of the caries from the sign (as starting point) to the deepest point of caries in the pictures and its length was recorded for a.The line a was lengthened to the contralateral edge of occlusal surface in the photo and the length was recorded for b.A line from the marked point on the occlusal surface edge of the extracted teeth was draw parallel to the line b on the corresponding photo and its length was recorded for c.The depth of the cavities on the projected images was recorded for d,and calculated d/a =c/b (digital optical fiber measured decay depth/caries damage depth of the image =actual tooth width/tooth width of the image),and d =c/b × a inferred.At last,the teeth were taken out from the standard model dentition.The decay of the tooth was removed completely.The actual depth of the cavity was recorded for D.The difference between d and D was recorded for Δd.The software of SPSS 20.0 was used to test the consistency of the results,and the MedCalc 14.8.1.0 software was used for Bland-Altman analysis.Results:The intraclass correlation coefficient (ICC) between d and D was 0.951 (ICC ＞ 75 ％),P =0.263.There was a function relationship y =0.23 ± 0.9 1x between d (x) and D (y).Bland-Altman analysis method showed that the mean of Δd (Δd) was 0.05 mm,the standard deviation of Δd (ΔdsD) =0.308,and the 95％ confidence interval was (-0.55 to 0.65).The amplitude of difference was clinically acceptable.So the consistency of the two measurement modes was high.Conclusion:There was no significant difference between the depth of caries lesions checked with DIAGNOcam system and the depth of the actual cavity,and the consistency was very good.The vitro study suggests that the DIAGNOcam system may be used to assess the depth of caries cavity as a useful tool in diagnosis and treatment.

Histocytochemistry ; methods ; Humans ; Pathology, Clinical ; methods ; Specimen Handling ; methods ; Staining and Labeling ; methods

OBJECTIVE:To study the technical conditions and parameters of DiaionHP-20 macroporous adsorption resin for the purification of the anti-PMOP(primary Type I osteoporosis) active fraction from Camellia semiserrata seed.METHODS: With the elution ratios and purity of total flavonoids and polyphenol from Camellia semiserrata as indexes,the technical parameters of DiaionHP-20 macroporous adsorption resin including the adsorption quantity,the loading sample,the elution concentration and volume of alcohol were investigated.RESULTS: The adsorption quantity of DiaionHP-20 macroporous resin on total flavonoids and polyphenol were 59.5 mg?g-1 and 23.9 mg?g-1 respectively.The optimal technical conditions were as follows:the quantity of drug was 10.5 times that of resin;water-soluble foreign substance was washed off with distilled water weighing 3 times the weight of resin,and total flavonoids and polyphenol were eluted using 25% ethanol weighing 13.5 times the weight of resin.CONCLUSION:DiaionHP-20 macroporous resin had good purification effects on total flavonoids and polyphenol from Camellia semiserrata seeds.The process is simple,low cost,and suitable for lavge-scale production.

Actins ; metabolism ; Adenoma, Pleomorphic ; congenital ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Fibrosarcoma ; metabolism ; pathology ; Humans ; Infant ; Male ; Nasopharyngeal Neoplasms ; congenital ; metabolism ; pathology ; surgery ; Rhabdomyosarcoma ; metabolism ; pathology ; Salivary Gland Neoplasms ; congenital ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Actins ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Back ; Desmin ; metabolism ; Diagnosis, Differential ; Fibrosarcoma ; pathology ; Humans ; Infant ; Male ; Neoplasm Recurrence, Local ; Reoperation ; Rhabdomyosarcoma ; drug therapy ; pathology ; surgery ; Vimentin ; metabolism

Objective To investigate the relationship between single nucleotide polymorphism (SNP) of Fas ligand (FasL) and fulminant hepatitis B in Han Chinese. Methods HBV infected subjects were enrolled in this case-control study, including 233 cases of inactive HBsAg carrier, 68 patients with fulminant hepatitis B,100 cases of spontaneous hepatitis B clearance, 102 patients with hepatitis B virus (HBV) related cirrhosis and 112 patients with HBV related primary hepatocellular carcinoma. The blood samples and clinical data were collected. FasL-844T/C polymorphisms of enrolled subjects were examined by TaqMan real time fluorescent genotyping polymerase chain reaction (RT-PCR). A adjusted odds ratios (OR)and 95% confidence intervals (CI)were calculated using the Logistic regression model. Results After adjusting the factors of gender and age, binary Logistic regression analyses indicated that the genotype frequencies of FasL-844 CC,CT,TT in inactive HBsAg carriers were 50. 64% ,39. 91% and 9. 44% respectively, and those in cases of fulminant hepatitis B were 79. 41%, 17. 65% and 2. 94%, respectively. The analysis also revealed that FasL-844CC genotype in inactive HBsAg carriers was high risk factor of developing fulminant hepatitis B (OR =4. 729,95%CI:0. 510 - 21. 282,P = 0. 043), while there were no statistic significances in other cases (P>0. 05). Conclusion The inactive HBsAg carriers harboring FasL-844CC may have greater susceptibility to fulminant hepatitis B, which need arouse high attention.