Aptazymes are a new artificial synzyme, selected from the random oligonucleotide sequence libraries against various of effector molecules. They own the advantages of an aptamer(the receptor site) and the ribozyme (the catalytic active site). Moreover, aptazymes as catalytic molecular beacon provide a new orientation for the quantitative analysis of effector molecules. Aptazymes not only have the application in genomics and proteomics, but also have potential applications in biosensor and DNA AND gate.

With the economic development, more and more people concerned about health. Institution and people engaged in physical examination is springing up. Facing increasingly less obvious advantages of product homogeneity, and the growing market which must cater to different customers, as a large general hospital medical center, we should make good management of customer relationship, improve customers' satisfaction, increase their loyalty, provide the most effective health protection for the physical examination so as to improve market competitiveness and bring social and economic benefits to the hospitals, thus achieving a win-win objective.

0.05).The smoking group had a higher TG level and a lower HDL-C level than the control one with statistical significance(P0.05).The HL activities in serum,lung and liver of smoking group were lower than those of control group(P

Objective To investigate the effect of ketamine injected via the radicular arteries on spinal cord. Methods Twenty healthy mongrel dogs of both sexes weighing 12-18 kg were randomly divided into 2 groups ( n = 10 each): control group (group C) and ketamine group (group K). The animals were anesthetized with intravenous pentobarbital 30-35 mg/kg, fentanyl 50-100 μg and vecuronium 0.2 mg/kg and maintained with propofol ically ventilated after tracheal intubation. A catheter was inserted into T8 poster intercostal artery and advanced toward the opening of radicular artery which supplies the spinal cord. Ketamine 100 mg (in 2 ml of normal saline)was injected via the catheter in group K. Three hours after ketamine administration, the animals were sacrificed. A 1.5 cm long segment of spinal cord at the level of T8 was removed for microscopic examination and determination of the expression of NSE, S100β and Tau protein by immuno-histochemistry. Results There was no significant difference in the number of Nissl' s staining-negative neuronal cells and the expression of NSE, S100β and Tau protein in the spinal cord between the 2 groups ( P ＞ 0.05 ). Conclusion Ketamine injected via the radicular arteries does not induce spinal cord injury.

To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 Vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from the total RNA of H9 cells. The resulting DNA construct was cloned into a prokaryotic expression vector (pET-32a). Recombinant pET-vif and pET-APOBEC3G were expressed respectively in Eserichia coli BL21 (DE3) as an insoluble protein. The vector also contained a six-histidine tag at the C-terminus for convenient purification and detection. To express and purify the HIV-1 Vif and hAPOBEC3G in E. coli cells, the accuracy of inserted gene and specificity of proteins were detected by the two enzyme digestion method, SDS-PAGE, and Western blotting. Rabbits were then immunized by Vif or APOBEC3G protein and serum samples were tested by indirect ELISA to determine the level of antibodies. Immunoenzyme and immunofluorescence assays were performed to identify the specificity of polyclonal antibodies. The titer of the anti-Vif antibodies was 1:204800, and that of the anti-APOBEC3G antibodies was 1:102400. Thus the antibodies could detect the antigen expression in the cells, demonstrating that fusion proteins with high purity and their corresponding polyclonal antibodies with high titer and specificity were achieved.

Aim To develop a rapid and inexpensive method for purification of human albumin, a method of immunomagnetic microspheres (IMMS) based on enzyme-linked immunosorbent assay (ELISA)for the purification of human albumin from human serum. Methods Polystyrene magnetic microspheres with carboxyl groups as carriers were prepared, and then the carboxyl groups on the surface of the microspheres were activated by ethylcarbodiimide (EDC). Finally rabbit anti-human serum albumin (HSA) antibodies were covalently bound to it and the complex can specifically capture HSA. After the procedure of capturing HSA, through taking rabbit anti-human albumin protein antibodies as a capture antibody, and goat anti-human albumin protein antibodies as a detection antibody, an ELISA on IMMS was developed, which can determine the recovery yield of HSA from the human serum. Results The result of the experiment was that the recovery of human albumin with IMMS was (86 ± 4) % , and IMMS were reused for two other purifying cycles, the results of which were (69.0 ± 0.6) % and (40.8 ± 0.8) % , and the purity of the product was about 90%. Conclusion The results above prove that the immunomagnetic purifiying strategy was shown to be efficient and offers an new thought for a large scale production of highpurity HSA.

ObjectiveTo establish a analytical system for the survival motor neuron (SMN) subtle mutation,and evaluate its application in two families with spinal muscular atrophy (SMA).MethodsSMN genes in seven family members from two SMA families were analyzed at both transcript level and genomic level,by the use of the conventional PCR-RFLP,allele-specific PCR,multiplex ligation-dependent probe amplification (MLPA) and T subcloning and sequencing of SMNI gene.ResultsIn family A,the patient had a single SMN1 copy who was carrying nonsense mutation L228X,which was also found in his father.In family B,as the patient's sample was unavailable,the father was indeed a carrier with one normal SMN1 allele and the other SMN1 allele carrying a frameshift mutation 22_23insA.The remaining family members were SMA carriers with one SMN1 copy.ConclusionThis analytical system for SMN subtle mutation offers viable molecular basis for genetic counseling and prenatal diagnosis in SMA families.

Objective To summarize the diagnosis and treatment experience of the pancreatic fistula secondary to abdominal operations.Methods The clinical data of 27 patients with pancreatic fistula due to abdominal operations were analyzed retrospectively.Results 25 patients were diagnosed by the amylase concentration of the drainage fluid and 2 patients were diagnosed by the percutaneous puncture fluid amylase concentration.Four patients underwent percutaneous puncture drainage by BS-guide.Five patients underwent re.operation drainage.Enteral feeding,total parenteral nutrition,total parenteral plus oral nutrition were applied to 15,6 and 6 patients,respectively.Altogether 3 patients died,all of these patients were in the total parenteral nutrition group.13 cases were discharged with draining tubes,including 2 patients who developed Dseudocyst and received surgical treatment,and the others 1 1 patients were discharged with tubes for(9.0±3.2)months.The mean hospital stays for oral feeding,jejunum tube nutrition and total parenteral nutrition groups were(36.3±10.2)d,(57.6±17.3)d and(63.3±33.4)d,respectively;and difference was statistically significant(F=3.49,P=0.049).The mean hospital stays for patients with or without somatostatin treatment were(53.5±20.3)d and(51.5 ±21.0)d,and difference was not statistically significant(t=0.207,P=0.838).Conclusions hereasingthe understanding ofpancreaticfistula,adequate drainage and rational nutrition phyed a key role in impmving the treatment effects of pancreatic fistula.

Background and purpose: High expression of telomerase and telomere stability are two common features in tumor cell. hTERT is a catalytic subunit of telomerase, TRF2 is extremely important to maintain the length and stability of telomerase. This study was to construct the recombinant adenovirus mediated shRNA to hTERT and TRF2, and to investigate the inhibitory effects of the vector by solo-inhibiting and connect-inhibiting in the MCF-7 cells, in order to present a new approach to the gene therapy for breast cancer. Methods: rAd-hTERT and rAd-TRF2 were constructed and the expression of hTERT mRNA and TRF2 mRNA were tested by FQ-PCR 48 hours after transfecting in MCF-7 cells. Apoptosis was observed by flow cytometry 1 to 6 days after transfection. Results: ①At 48 hours after transfection, the results of FQ-PCR showed that compared to PBS group, the expression of hTERT in rAd-hTERT group was obviously decreased and the inhibition ratio was about 86%, but TRF2 had not been obviously inhibited (P>0.05);the expression of TRF2 in rAd-TRF2 group was obviously decreased and the inhibition ratio was about 80%, but hTERT had not been obviously inhibited (P>0.05);in rAd-hTERT/rAd-TRF2 group, the inhibition ratio of hTERT and TRF2 were about 88% and 85%. Comparing rAd-hTERT/rAd-TRF2 group with rAd-hTERT group and rAd-TRF2 group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). Otherwise, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). ②The result of flow cytometry showed that apoptosis was induced at the first day after transfecting in rAd-hTERT group and rAd-TRF2 group, the most obvious apoptosis was in the 3rd to 5th days,at the peak in the 5th day, and decreased in the 6th day after transfection. The apoptosis ratio of rAd-hTERT group was 46.2%, rAd-TRF2 group was 43.5%. The apoptosis ratio of rAd-hTERT/rAd-TRF2 group was 46.2% at first day, 68.5% at the second day, the most obvious apoptosis was in the 3rd to 6th days and was 77.6% in the 6th days in rAd-hTERT/rAd-TRF2 group. There were significant differences in apoptosis ratio in solo-inhibiting and connect-inhibiting(P<0.05). In addition, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences in apoptosis ratio(P>0.05). Conclusion: ①Target sequence of RNAi which aimed at hTERT gene and TRF2 gene was designed efficiently, and the RNAi expression vectors were seen in vivo study efficiently and specifically inhibited the correspond gene expression and promoted cell apoptosis in MCF-7 cells. ②rAd-hTERT vector and rAd-TRF2 vector have no synergistical effect and antagoinstical effect on inhibiting hTERT gene and TRF2 gene mRNA expressing in MCF-7, but there was synergistical effect in terms of the induction of apoptosis. So association-RNAi-technique targeting to the genes of telomere length and stability can effectively promote tumor cell into apoptosis and inhibit breast cancer cell growth. RNAi technique of connecting correlation genes is a more effective gene therapy strategy.

Humans ; Stomatitis, Aphthous ; diagnosis ; etiology ; therapy