Objective To investigate the correlation of interleukin-1 family genotypes,including interleukin-1 (IL-1α,IL-1β) and interleukin-1 receptor antagonist (IL-1Ra),with coronary heart disease (CHD) and serum lipoprotein level in the elderly.Methods Interleukin-1 family genotypes were detected in 318 elderly controls and 329 elderly CHD patients by polymerase chain reaction and restriction fragment length polymorphisms method.Serum levels of lipoproteins were inspected simultaneously.Results The TT and Ⅰ / Ⅰ or Ⅰ/Ⅳ genotype frequency of IL-1Ra was 90.3％ in elderly CHD patients,but 82.4％ in controls.Carriers with TT,Ⅰ / Ⅰ or Ⅰ/Ⅳ genotype of IL-1Ra were at an increased risk with an odds ratio of 1.98 in elderly CHD patients as compared with controls (x2=8.55,95％ CI:1.25-3.16).The TT and Ⅰ/Ⅰ or Ⅰ/Ⅳ genotype frequency of IL-1Ra was 96.2％ in elderly CHD patients with acute coronary syndrome,but 84.8％ in elderly CHD patients with stable angina.Carriers with TT,Ⅰ / Ⅰ or Ⅰ/Ⅳ genotype of IL-1Ra were at an increased risk with an odds ratio of 4.54 in acute coronary syndrome group as compared with stable angina group (x2=12.17,95％CI:1.81-11.36).The CT or TT genotype frequency of IL-1α-889 was 22.8％ in acute coronary syndrome group,but 7.6 ％ in stable angina group.Carriers with CT or TT genotype of IL-1α-889 were at an increased risk with an odds ratio of 3.59 as compared with stable angina group (x2 =14.93,95％CI:1.82-7.03).There were no significant differences in levels of serum lipoproteins among the different genotypes (P＞0.05).Conclusions In elderly patients with coronary heart disease,IL-1α(-889) CT or TT genotype carriers are at high risk for acute coronary syndrome,but IL-1Ra CC,TC,Ⅰ / Ⅱ or Ⅱ / Ⅱ genotype carriers are at a low risk for CHD or severe CHD.

Objective To investigate the change of serum homocysteine (Hcy) and plasma fibrinogen (Fg) levels in patients with ischemic cerebrovascular diseases (ICVD) and the correlation between Hcy and Fg.Methods Blood samples were collected from 74 patients with ICVD and 40 controls. The concentrations of Hcy, folate and Fg were determinated by means of fluorescence polarization immunoassay, electrochemiluminescence, and immunoturbidimetry, respectively. In the meantime, plasma lipid was detected by routine method.Results Both Hcy and Fg levels in the patients with ICVD increased significantly as compared with the controls (all P0.05). Multivariate logistic regression analysis indicated that hypertension, high cholesterol, high Fg and high Hcy concentrations played significiant roles in ICVD.Conclusion Both hyperhomocysteinemia and hyperfibrinogenemia are independent risk factors of ICVD.

Objective: To establish the callus culturing system of embryos in Sophora japonica, and to analyze the contents of isoflavones in callus. Methods: Through adding the plant growth regulators of different kinds and at different concentration, the optimal media for callus induction, suspension cell culture, and the highest isoflavone content were obtained. Results: The optimal medium for callus induction of embryos in S. japonica was B5 + 2, 4-D (1.0 mg/L) + 6-BA (0.2 mg/L) + sucrose (20 mg/L). In solid medium, the callus was growing rapidly after 7 d, and the yield reached to the highest on the day 35 (fresh weight 7.4137 g), but the isoflavone content had no significant changes. In suspension culture, the yield of callus reached to the highest on the day 24 (fresh weight 11.563 8 g), and the callus turned into decline phase after 24 d, in which the fresh and dry callus weights were descending. With continuous culture, the cells were browning gradually, aging, and dying at last, but the isoflavone content was opposite to growth curve, and reached its lowest point (4.826 mg/g) on the day 24. Conclusion: The induction and cultivation of embryos callus in S. japonica and the isoflavone content have a larger correlation with the plant growth regulators of the kinds and the concentration.

To construct a recombinant adenovirus containing CDglyES fusion gene, which can directly inhibit human ovarian cancer cell and indirectly inhibit vascular endothelial cell growth. Methods:We constructed prAdCDglyES using a homolo-gous recombination method in bacteria. The prAdCDglyES was transfected to 293 packaging cells using liposome, in which rAdCDgly-ES was packaged and amplified. MTT was used to observe the proliferation inhibition effect of rAdCDglyES on human ovarian cancer cells and the growth inhibition effect of expressing products of rAdCDglyES on ECV-304. Results:The titer of rAdCDglyES was 1 × 1013.3 TCID50/L, whereas the inhibition rate on human ovarian cancer cell SKOV-3 was (83.1±6.3)%. This result is significantly different from the control rAd-LacZ, which had an inhibition rate of (24.1 ± 13.2)% (P<0.01). The concentrated culture supernatant from cells transfected with rAdCDglyES can inhibit ECV-304 cell proliferation at a rate of (78.7 ± 1.6)%. This rate is significantly different com-pared with that of the control with the same concentration of culture supernatant from cells transfected with rAd-CD, with an effect on ECV-304 cell shown by an inhibition rate of (23.9 ± 9.7)%(P<0.01). Conclusion:The results showed that the recombinant adenovirus rAdCDglyES could inhibit human ovarian cancer cells directly and indirectly.

Objective:To generate rabbit polyclonal antibody against human Argonaute2 (Ago2) protein and to identify its functional characterization for determination of differential expression and cellular localization of Ago2 protein in various cell lines.Methods:DNAstar software was applied for searching the high antigenicity region of Ago2 gene sequence termed k-Ago2.Prokaryotic expressing plasmid was constructed and transformed to E.coli BL21 (DE3) to induce expression by IPTG.The fusion protein was injected into rabbits subcutaneously to produce polyclonal antibodies after purification by gel regaining.ELISA was operated to detect antibody titer.Western blot was used to identify the specificity and sensitivity of the antibodies and detect the differential expression of Ago2 protein in various cell lines.Meanwhile,immunofluorescence experiments were arranged to show cellular localization of Ago2 protein.Results:The prokaryotic expressing plasmid was constructed correctly.K-Ago2 protein was expressed and purified,and then rabbit polyclonal antibodies against Ago2 were generated after immunization with k-Ago2 protein.The titer detected by ELISA was 1∶19 000.Western blot results demonstrated the high specificity of the antibodies.Finally,we successfully observed the differential expression and cellular localization of Ago2 protein in various cell lines.Conclusion:The polyclonal antibody against Ago2 protein has been achieved successfully.It will be propitious for the intensive study of the RNAi mechanism and even profound clinical application.

Objective To evaluate the embolic effect on rabbits arteries using self-made copper plated platinum coil.Methods 1 7 New Zealand Big Ear Rabbits were selected.Unilateral subclavian artery or carotid artery was embolized with self-made secondary level copper plated platinum micro-coils (experimental group)through 3F-catheter.Contralateral subclavian artery or carotid artery was also embolized using secondary level platinum micro-coils (control group)as a control.The level of serum copper ions and the liver and renal function were recorded during different intervals before and after embolization.The arteriography and the tissue his-tology were observed respectively during different intervals after the embolization.Results 1 5 of 1 7 rabbits were embolized success-fully.After embolization,the level of serum copper ions increased in 2 weeks(P <0.05).However,it returned to preoperative level after 4 weeks (P >0.05).The liver and renal function was similar to that of the preoperation after 2 weeks.After embolization,an-giography showed that vascular embolization effect between two groups was not significantly different at 10 min and 30 min;howev-er,the embolization effect of experimental group was superior to that of control one (P <0.01)at 3 days and 1 week,2 weeks,4 weeks,6 weeks and 12 weeks.Pathological results showed that there were a lot of thromboses inside,outside and around the copper coil.Few thromboses appeared around platinum coil in control group.The thrombosis situation in experiment group was better than that in control one (P <0.01).However,no significant difference in inflammatory cell infiltration between two groups was found. Conclusion Self-made secondary level copper plated platinum coil has good physical property,rememorability,flexibility and con-trollability.

Forearm ; innervation ; Ganglion Cysts ; surgery ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; innervation ; surgery ; Tendons ; surgery ; Ulnar Nerve ; surgery

Objective To establish a method of gold nanoprobe-based solution hybridization (GNBSH) to detect nucleic acid sequence-based amplification (NASBA) products for the rapid diagnosis of invasive aspergillosis (IA).Methods The Aspergillus specific 18S rRNA was amplified by NASBA and then the amplified products were hybridized with the gold nanoprobes which were modified with thiol compounds at the 5'end.Serum samples from 106 patients,including 14 with a definite IA,32 with suspected IA and 60 without IA,were detected by the established method,and the obtained results were compared with that of galactomannan (GM) test to evaluate its accuracy.Results The gold nanoprobes only hybridized with Aspergillus NASBA products but not other non-Aspergillus strains.The sensitivity,specificity and the area under the ROC curve (AUCROC) of the established GNBSH method for detecting 106 clinical samples were 82.61％ (38/46),81.67％ (49/60) and 0.890,respectively.The sensitivity,specificity and AUCROC of GM test were 56.52％ (26/46),83.33％ (50/60) and 0.723,respectively.Conclusion The established GNBSH method to detect Aspergillus NASBA products has high sensitivity and specificity and simple operation,which may be used to detect the infection of Aspergillus by clinical laboratories.

Objective To investigate the dose- and time-effects of astragaloside Ⅳ(XGA) on collagen of myocardial fibroblasts in rats.Methods The myocardial fibroblasts of rats were separated by collagenase and trypsinase digestive method,and the cell culture system was established. After XGA in different concentrations and at different time points was administered in fibroblast culture systems,the mRNA expression levels of collagen,matrix metalloproteinases(MMP)-1,-2,-9,tissue inhibitor of metalloproteinase(TIMP)-1 and -2 were measured with reverse transcription-polymerase chain reaction(RT-PCR) test.Results After XGA administration with different doses and at different time points was adminstered,the gel electrophoresis product of RT-PCR in fibroblast culture system expressed the mRNA of type Ⅰ,Ⅲ and Ⅳ collagens,MMP-1,-2,-9,TIMP-1 and -2;but the mRNA expression levels of type Ⅰ,Ⅲ and Ⅳ collagens,TIMP-1 and -2 decreased and the mRNA expression levels of MMP-1,-2,and -9 increased compared to those before XGA administration;the mRNA expression of type Ⅰ,Ⅲ and Ⅳ collagens,MMP-1,-2,-9,TIMP-1 and -2 decreased or increased gradually with the increase of doses and the prolonged time of XGA use.The mRNA expression levels of type Ⅰ,Ⅲ and Ⅳ collagens,TIMP-1 and -2 were negatively related to the doses and action times of XGA(r=-0.927 to -0.637 P= 0 to 0.024);and the mRNA expression levels of MMP-1,-2 and -9 were positively related to the doses and action times of XGA(r=0.672 to 0.962 P=0 to 0.034).Conclusion XGA can markedly reduce the collagen formation of myocardial fibroblasts in rats,and its mechanisms are related to the inhibiting of collagen synthesis and the increase of collagen degradation.

Aim To investigate the effects of 3 ,5 ,2 ’ , 4’-tetrahydroxychalcone (P40) on urate excretion, as well as mRNA and protein expressions of renal URAT1 and GLUT9 in hyperuricemic mice. Methods Sixty Kunming mice were randomly divided into six groups:normal control group, hyperuricemic group ( model group), P40 2. 0, 4. 0, 8. 0 mg·kg-1 groups and positive control group. All drugs were administered in-tragastrically to mice for 5 doses. Hyperuricemic mice were induced by intraperitoneal injection of uric acid (0. 15 g·kg-1 body weight) for 3 times. The urate levels were assayed with the phosphotungstic acid method. The mRNA and protein expressions of GLUT9 and URAT1 were determined by RT-PCR and Western blot. Results P40 at a dose of 4. 0 and 8. 0 mg · kg-1 significantly reduced the serum urate levels in a dose-dependent manner, when compared with untreat-ed hyperuricemic mice ( P<0. 05 or 0. 01 ) . The he-patic urate contents decreased in untreated-and treated-hyperuricemic mice as compared with normal mice ( P<0. 01 ) . Furthermore, P40 had no influence on the renal URAT1 mRNA and protein expression levels, while it could down-regulate renal GLUT9 protein ex-pression but not mRNA expression in hyperuricemic mice. Conclusion P40 possesses potent uricosuric effects associated with urate reabsorption by down-regu-lating the protein expression of GLUT9 in kidney.