Cancer stem cells are considered to be the origin of tumor relapse.Isolating and identifying them by special immune-phenotypes is the most commonly used method.The technique of isolating SP cells is very mature.More attentions are being paid to the isolation of cancer stem cells using differential expression activities of Wnt pathway on surface of tumor cells.Establishing stem cells niches to capture cancer tumor cells remains to be further evaluated.

Neonatal acute heart failure is a common critical illness,and also one of the material cause of perinatal death. It is difficult for early diagnosis due to the different characteristics and clinical manifestation between neonate and older children, which leads to the difficulty of diagnosis timely and affect the condition adversely. This article introduced the common etiology, characteristics of clinical manifestation, diagnosis and treatment of neonatal acute heart failure.

Objective To investigate the clinical application of Cystatin C as a biological marker for monitoring hepatic pathological change in patients with virus hepatitis.Methods Two hundred and seven patients infected with hepatitis B or C virus(HBV, HCV)were divided into cirrhosis group(group A),chronic HBV group(group B),chronic HCV group(group C),and liver cancer group(group D). 32 healthy controls(group H) were recruited . The serum TIMP-1,TIMP-2,and Cystatin C as well as some traditional markers for monitoring liver function and renal function including creatinine, creatinine clearance rate, alanine transaminase, and aspartate transaminase were determined.Results In these groups, serum Cystatin C(F=28.334, P

The cell suspensions prepared from surgically resected hepatoma specimens were used to immunize BALB/c mice and, with the hybridoma technique, a battery of high affinity monoclonal anti-hepatoma antibodies were obtained which were designated HAbl8, Fll, E5 and A10 separately. Immunohistochimical staining showed that the above 4 antibodies possessed good selective reactivity with hepatoma tissue. After radioiodination of Fll, E5, A10, HAbl8 IgG and It's F(ab')_(2) fragments, the labelled reagents were employed for radioimmunoimaging in hepatoma-bearing nude mice and the in vivo detection appeared promising, with tumor/non-tumor ratios being 6.88, 5.14, 5.67, 5.15 and 14.47 respectively. The in vivo localization ablities of the antibodies seemed better compared to other similar findings published elswhere (Dunk AA, 1987). Also, ~(131) I -HAbl8 I gG and its radiolabelled fragments were utilized for radioimmunotherapy in hepatoma-bearing nude mice, with complete response rate and partial response rate being 42%(5/12) and 50% (6/12)respectively. When the HAbl8 conjugate with radioiodine was introduced for the in vivo imaging in hepatoma patients, a positivity rate of 86.5% (45/52)was witnessed, with the smallest tumor foci detected being only 0.5cm in diameter. In the in vivo targeting therapy with the immunoconjugate, a partial response rate of 69.6% (16/23) was obtained. In summary, the antibodies reported here have lended a novel regime for the present comprehensive therapy protocol of hepatoma.

Objective:To prepare monoclonal antibodies(McAb)against cystatin C(Cys C)and to establish the particle enhanced turbidimetric immunoassay(PETIA)for determining human serum Cys C.Methods:The prokaryotic expression vector pET32a(+)/Cys C was constructed and Cys C expression was induced.McAbs against Cys C were prepared with the hybridoma technique after mice were immunized with the purified recombinant protein.Then the McAbs were covalently attached to uniform microparticles,PETIA method for determination of human serum Cys C was established,and primary evaluation tests of methodology were performed.Results:Three hybridoma cell lines were obtained successfully,the secreted antibodies were isotype of IgG1,and Western blot confirmed that the antibodies reacted specifically to the Cys C protein.After one of the hybridoma cell lines was injected into mice abdominal cavity,the ascites abundant for McAb was obtained.The titer of the McAb against the purified protein was 1∶4?106.With the self-made McAb,PETIA for human serum Cys C was established.The primary evaluation tests of methodology revealed that self-established PETIA method had a satisfactory performance,which was equal to the import kit.Conclusion:The prepared McAb against Cys C is prepared,which could be used to establish PETIA for determining human serum Cys C.

Objective To investigate the relationship between angiotensin I-converting enzyme(ACE)inserting/defaulting(I/D) gene polymorphisms and the femoral artery intima-media thickness(FA-IMT) in patients with type 2 diabetes mellitus(T2DM).Methods The polymorphisms of ACE(I/D) was determined by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method and the FA-IMT was assessed using non-invasive high resolution B-mode ultrasonography in 303 patients with T2DM in Hunan province.Results The frequency of I allele of ACE gene polymorphisms was higher in T2DM than that in healthy controls,but frequency of D allele was lower in T2DM than that in healthy controls(P

Objective: To investigate the value of enhanced 3D SPACE sequence in displaying brachial plexus. Methods:35 healthy volunteers with no history of brachial plexus injury of brachial plexus MRI examinations by DWIBS scan, and SPACE scanned. Analysis and comparison of DWIBS sequences, sequences of SPACE provides a clear display of the brachial plexus, and two sets of image sequence average signal-to-noise ratio (SNR) and the contrast to noise ratio (CNR). Results:A total of 35 cases 70 lateral brachial plexus after coronary DWIBS is clearly displayed on the sequence;the partial medial cord of brachial plexus, beams and lateral beams can also be displayed in the SPACE sequence. SPACE sequence of image signal-to-noise ratio and contrast noise ratio than the DWIBS sequence, and the difference is statistically significant. Conclusion:SPACE sequence can clearly show the brachial plexus, and DWIBS sequences compared to the normal image has a higher resolution of the brachial plexus.

AIM To establish the method for the enzyme assay of dimethylarginine dimethylaminohydrolase (DDAH) in rabbit kidney and to determine the optimal condition for the assaying. METHODS Five healthy Japanese rabbits weighing 3 0 to 3 5 Kg were killed by air embolism,kidneys were harvested and then were homogenized. Asymmetric dimethylarginine (ADMA) was used as the substrate for DDAH. UV 265 spectrophotometer was applied to determine the amount of the enzymatic product L Citrulline( L Cit).The amount of L Cit produced under different conditions was compared and the optimal condition was screened. The kinetic parameters of ADMA degraded by DDAH were calculated. RESULTS The kinetic parameters of ADMA degraded by DDAH were as follows: K m=(0 28?0 10) mmol?L -1 , V m=(1 36?0 42) mmol?L -1 ?g -1 ?min -1 . The optimal conditions for the enzyme assay of DDAH in rabbit kidney determined in this study follow: The concentration of the enzyme protein was 12 g?L -1 ,the optimal pH of the buffer was 7 4,the final concentration of ADMA was 2 mmol?L -1 ,and the reaction time was 30 min. CONCLUSION The method offered here is easily done. The concentration of the substrate determined in this study was based on the value of Km,thus it was beneficial to the accurate assay of DDAH.

Objective To construct a prokaryotic expression vector of cystatin C (Cys C), purify Cys C protein produced by the expression system, and prepare its antiserum. Methods Total RNA was isolated from HL-60 cells, and human Cys C gene was amplified with RT-PCR. The cDNA fragment was cloned into pMD18-T vector and which was confirmed by sequencing. The enzyme-digested target fragment was cloned into PET-32(a) expression vector and transfected into E.coli. BL 21(DE3), in which Cys C expression was induced. After the inclusion body protein was purified through Ni2+ affinity chromatography, processed by dialysis, identified by Western blotting, a rabbit was immunized with the fusion protein, and the antiserum was obtained. Results The result of DNA sequence analysis showed that the cloned Cys C gene sequence was completely corresponding to GenBank data. SDS-PAGE and Western blotting showed that the expressed Cys C fusion protein was about 35?103, mainly existing in the inclusion body of E.coli., that could be purified through Ni2+ affinity chromatography. The titer of the antiserum to the purified protein was 1∶8 000 by ELISA, and Western blotting confirmed that the antiserum reacted specifically to the Cys C protein. Conclusion A recombinant Cys C protein and the specific polyclonal antibody have been obtained, which provides a basis for establishment of immunoassays of human Cys C.

Objective To establish a continuous monitoring assay of serum argininosuccinate lyase (ASL) activity with automatic biochemistry analyzer, and perform methodology validation and preliminary clinical application.Methods According to the chemical reaction catalyzed by ASL and the working characteristics of automatic biochemistry analyzer, an enzyme coupled reaction system with high specificity was set up, and the methodology validation was performed.Three hundred and nine patients with various liver diseases, 269 non-liver disease patients and 40 healthy controls were enrolled in this study.Serum ASL, ALT, and AST level were determined in all subjects.Results A new kinetics assay of ASL activity was set up with automatic biochemistry analyzer.The methodological validation demonstratod that inter-assay and intra-assay coefficient of variation were 4.0% and 5.9% respectively and the mean recovery was 100.5%.The linear range was 0-167.7 U/L.The lowest detection limit was approximately 0 U/L.The interference test showed that there is no significant interferences while the concentration of bilirubin is less than 342 μmoL/L or commonly used anticoagulants is employed at their routine concentrations.However,interference was significant when Hb level is more than 0.06 g/L.Preliminary study of clinical application showed that there was no significant difference of serum ASL level between non-liver disease group and healthy group ( q = 0.027, P = 0.979 ), but there was significant differences for both serum ALT and AST levels (ALT:q =6.461,P =0.000;AST:q =6.481,P =0.000).Conclusions A continuous monitoring assay for the determination of serum ASL activity is successfully established. Serum ASL may be a good biomarker for liver injury.