Objective To analyze the factors of Protein A/G affinity column,influencing purification effect,and get the best condition to improve application effect of Protein A/G affinity column. Methods We Purified anti-HCV-IgG serum with different disposal modality,different sample input modality and different application number of affinity column before detecting and analyzing the purified samples. Results The Protein A/G affinity column had the best purified effect after using saturated ammonium sulfate to first purification,which increased the affinity column adsorption effect within 30 minutes adsorption. Conclusion Using antibody with first purification and adding the adsorption time could improve utilization rate of affinity column and prolong the service life.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Chromatography, Gas ; methods ; Hydrocarbons, Chlorinated ; analysis

OBJECTIVE:To explore the relevant factors that led to the adverse drug reaction(ADR)of netilmicin.METHODS:ADR cases resulted from netilmicin reported in12kinds of Chinese periodicals from1998to2003were analyzed statistically.RESULTS:The ADR of netilmicin involved multiple organs and systems,the clinical manifestation were compli-cated and diversified,which manifested mainly on the renal toxicity,toxicity of nervous system and allergic reaction.CON-CLUSION:Clinician and pharmacists should be alert to the ADR of netilmicin and keep using drugs rationally.

Objective To establish a method for determination of total hardness in drinking water by using automatic potentiometric titrator.Methods Dynamic equivalence point titration(DET)and monotonic equivalence point titration(MET)mode was used to determine the high total hardness and low total hardness of drinking water samples respectively.Results Used DET mode to determine the high total hardness,the relative standard deviation(RSD)was 0.69%～1.72% and the recovery rate was 101.5%～102.2%.Used MET mode to determine the low total hardness,RSD was 3.49%～4.00% and the recovery rate was 95.8%- 103.6%.Conclusion This method is rapid,simple,accurate,precise and applicable to the determination of total hardness in drinking water in low and high levels.

ObjectiveTo investigate the effect of early electro-acupuncture treatment on prevention of post-operative adhesions.Methods40 rabbits were randomly divided into 5 groups: normal control group, model group 1, model group 2, early electro-acupuncture group and later electro-acupuncture group with 8 animals in each group. Muscle adhesion model was established by operation. The early electro-acupuncture group was received the treatment from the 11th to 20th day after the operation; later electro-acupuncture group was from the 21st to 30th day after the operation. The weight, gait and range of motion (ROM) of animals were observed and analyzed.ResultsThe standing, walking and ROM of animals of the early electro-acupuncture group improved; but only few gait indexes improved and ROM not changed in the later electro-acupuncture group.ConclusionThe early electro-acupuncture treatment can prevent the post-operative adhesion of muscles, but not having effect on formed adhesion.

Objective To investigate effect of different doses of rosuvastatin on brachial artery endothelium-dependent vasodilation and carotid intima-media thickness in patients with coronary heart disease.Methods 92 patients with coronary heart disease in our hospital were admitted and divided into four groups according to randomly digital method, including 23 cases in control group were treated with lipid nitrate, antiplatelet aggregation, anticoagulant, lowering blood sugar, blood pressure control and other of conventional therapy;23 cases in group A, on the basis of conventional therapy, were treated with rosuvastatin 5 mg/d, orally, once daily;23 cases in group B were treated with rosuvastatin 10 mg/d, orally, once daily based on the conventional therapy;23 cases in group C were treated with rosuvastatin 20 mg/d, orally, once daily based on conventional treatment, each group was treated for 8 weeks.Brachial artery endothelium-dependent vasodilation (FMD) and carotid intima-media thickness (IMT) of patients before and after treatment were collected by color ultrasonic doppler, while observed lipid levels changes of 4 groups.Results Control group was treated for eight weeks, FMD, ITM, blood lipid levels and each index values were not significantly changed, the difference was not statistically significant;After treatment, total cholesterol ( TC) , low-density lipoprotein cholesterol C ( LDL-C) of A, B, C groups were significantly better than that before treatment, the difference was statistically significant (P<0.05), and decrease amplitude with dose of rosuvastatin increased became grearer, but the total cholesterol (TC), high density lipoprotein cholesterol C( HDL-C) there was no significant difference compared with before treatment; Compared with before treatment, ITM of A, B, C groups decreased, and the difference was statistically significant (P<0.05), decrease amplitude with dose of rosuvastatin increased became greater.Conclusion Rosuvastatin can significantly improve brachial artery endothelium-dependent vasodilation and carotid intima-media thickness in patients with coronary heart disease, and there is a clear dose-response relationship, which may be associated with rosuvastatin decrease total cholesterol and low-density lipoprotein cholesterol C in patients with coronary heart disease.It has guide significance to clinical.

AIM: To investigate lipoprotein lipase gene HindⅢ polymorphism and its relationship with serum lipids and apolipoprotein, serum HDL subclasses in patients with hyperlipoidemia. METHODS: Lipoprotein lipase gene HindⅢ polymorphism was assayed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The subclasses of serum HDL in 152 hyperlipoidemia patients and 128 healthy subjects were determined by two-dimensional gel electrophoresis conjunction with immunodetection method. RESULTS: H+H+ genotype and allele H+ in hyperlipoidemia and control groups were both the highest. In hyperlipidemia group, H+H+ genotypes tended to be higher than that in control group, while H+H- and H-H- genotypes were significantly lower (P

The new generation cyclooxygenase (COX-2) inhibitor could reduce the gastrointestinal side effect of NSAID drugs, but eventually increase the cardiovascular risk, because its selective inhibition of COX-2 induces the imbalance between PGI2 and TXA2 and the reduction of vasodilatory NO. Under pathological conditions, active oxygen species (O2-*2, etc) were used to induce endo- thelial dysfunction, activate NF-κB to induce expressions of pro-inflammatory cytokines IL-1β and TNF-α, increase ET-1, TXA2 with vasoconstrictor effect, reduce PGI2 and NO with vasodilatory effect, generate further oxidative damage together with NO, and reduce the bioavailability of NO. NO-NSAIDs and NO-Coxibs drugs raised the level of NO by introducing NO-donor (ONO2). NSAIDs drugs enhanced the anti-inflammatory activity of COX-2 and reduced gastrointestinal side effects by inhibiting selectively COX-2. If antioxidant structures with active ingredients of traditional Chinese medicines were introduced to improve the antioxidant activity of NSAIDs, they could scavenge the active oxygen species to protect the normal function of vascular endothelia and enhance the bioavailability of NO, which is conducive to enhance the cardiovascular safety of cyclooxygenase (COX-2) inhibitor.
Anti-Inflammatory Agents ; therapeutic use ; Biomarkers, Pharmacological ; Cardiovascular Diseases ; drug therapy ; enzymology ; immunology ; Cyclooxygenase 2 ; immunology ; Cyclooxygenase 2 Inhibitors ; adverse effects ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; NF-kappa B ; immunology ; Reactive Oxygen Species ; immunology ; Tumor Necrosis Factor-alpha ; immunology

AIM:To investigate the effect of calcitonin gene-related peptide (CGRP)transfection into c-kitpos cardiac stem cells (c-kit+CSCs) on the cell viability.METHODS: Under the sterile condition, the auricles of SD rats were taken out , and then c-kit +CSCs were collected through enzyme digestion and immunomagnetic bead separation (MACS).The cells were identified by flow cytometry .c-kit +CSCs were transfected with enhanced green fluorescent pro-tein CGRP lentiviral vector ( Lv-EGFP-CGRP) or enhanced green fluorescent protein lentiviral vector ( Lv-EGFP).The cells were randomly divided into Lv-EGFP-CGRP-CSCs group , Lv-EGFP-CSCs group and CSCs group .The transfection was observed under the fluorescence microscope .The transfection efficiency was detected by flow cytometry .The CGRP protein secretion in the cell culture supernatants was detected by ELISA .The viability of c-kit+CSCs transfected with Lv-EGFP-CGRP or Lv-EGFP was measured by CCK-8 assay.RESULTS:c-kit+CSCs were isolated and cultured successfully .The expression positive rate of c-kit was 91.0%and the expression positive rates of CD 45 and CD34 were 4.5% and 4.0%, respectively.After transfected with lentivirus for 48 h, the stable fluorescence in c-kit +CSCs was observed under fluores-cence microscope .The transfection efficiency were 80%when MOI was 20.The level of CGRP was significantly increased in Lv-ECFP-CGRP-CSCs group compared with Lv-EGFP-CSCs group and CSCs group (P<0.05).Meanwhile, transfection with lentiviral vector in each group did not affect the viability of c-kit+CSCs.CONCLUSION:Transfection of Lv-EGFP-CGRP into c-kit+CSCs was successful .The secretion of CGRP was found in the transfected c-kit+CSCs and the viability was not changed after transfection .CGRP-modified c-kit+CSCs may play a role in treating myocardial infarction .

Background Tight junctions are thought to play a significant role in the maintenance of the corneal epithelial defense for the eye,and the restoration of the tight junctions is critical during epithelial wound healing after refractive surgery.However,there are few reports about this study. Objective The aim of this study was to investigate and compare the expression of occludin in corneal epithelium following flap-free epipolis laser insitu keratomileusis (Epi-LASIK) and laser-assisted subepithelial keratectomy ( LASEK ). Methods Forty-eight clean New Zealand white rabbits were randomized into 2 groups,and 24 rabbits for each group.Flap-free Epi-LASIK and LASEK were performed in the right eyes of the rabbits in two groups,and other 2 age matched normal rabbits were as normal controls.The animals were sacrificed and the corneal samples were obtained at 1,2,3,5 days after surgery.Immunofluorescence staining was used to detect the expression of occludin in the corneal epithelium,and RT-PCR was used to identify the level of occludin mRNA in the central cornea. Results Occludin protein was expressed innormal corneal epithelium and showed the green fluorescence with the regular arrangement.The fluorescence intensity was lower in 1 -2 days in LASEK group with the irregular arrangement;while the fluorescence signal in corneal epithelium was stronger in flap-free Epi-LASIK group.3-5 days after surgery,the fluorescence intensities were bothenhanced in two groups.RT-PCR showed that the relative expression of occludin mRNA in corneal epithelium was 0.11 ±0.02,0.25 ± 0.03,0.43 ± 0.04 in LASEK group 1,2,3 days after surgery,respectively,and was lower than those of flap-free Epi-LASIK group ( 0.20± 0.04,0.44 ± 0.04,0.76 ± 0.04 ),showing significant differences between these two groups ( t =6.476,12.898,17.315,P＜ 0.05 ).No significant difference was seen in the expression of occludin mRNA in 5 days after surgery between two groups( t=-0.733,P＞0.05).The relative values of occludin mRNA expression in corneal epithelium were gradually increased with time prolongation,presenting a significant difference among various time points ( Ftime =768.903,P =0.000). Conclusions The reformation of occludin in flap-free Epi-LASIK group is faster than that in LASEK group.Therefore,flap-free Epi-LASIK is prominant in reducing the stimulated symptoms and complication after the surgery.