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Objective To investigate the stability of 8 commonly used reference solutions and determine the validity period of internal control. Methods The storage solutions of reference substances were prepared and stored in the refrigerator at 2 to 10 ℃. The content of the storage solutions and the newly prepared reference solutions were determined by HPLC on days 0, 1, 4, 7, 10, 14, 21, 28 and 35, and their change values of the content were calculated. Results During the inspection period, the appearance of each reference solution was consistent with the newly prepared reference solution. There was no significant impurity peak in the chromatography. For mixed references of metronidazole, chloramphenicol and salicylic acid stored for 7 days, vitamin E, mixed references of tinidazole and chlorhexidine acetate, sulfadiazine, dexamethasone phosphate for 35 days, their contents met the requirements. Conclusion Stored in the refrigerator at 2 to 10 ℃, the effective time period of vitamin E reference solution, mixed references solution of tinidazole and chlorhexidine acetate, sulfadiazine reference, dexamethasone phosphate reference solution can be 35 days, and the mixed references solution of metronidazole, chloramphenicol and salicylic acid defined can be 7 days.
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In recent years, the IVD industry has developed rapidly based on the increasing market demand, and plays an important role in disease prevention, clinical diagnosis, health monitoring and guiding treatment. Therefore, followed quality and safety issues are highly concerned. The unique advantages of blockchain technology, decentralization, distrust and non-tampering, can write into trusted node data in every link covering production, circulation and usage of IVD reagents, and establish a distributed ledger with full backup, which makes the anti-conterfeiting and traceability for IVD reagents possible. We discuss whole process intelligent tracing system for IVD reagents based on blockchain technology. Through the strong mechanism of pre-supervision and post-punishment, the source of reagents can be traced, quality and responsibility can be investigated, and the medical inspection quality and diagnostic safety can be guarded.
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Blockchain , Indicators and Reagents , Reagent Kits, Diagnostic , TechnologyABSTRACT
Objective To analyze the status of mycoplasma infection and drug resistance in the local area,and provide the ba-sis for clinical rational drug use.Methods The specimens obtained from 1 9 530 patients with urogenital tract infection were detec-ted by adopting mycoplasma culture,identification and drug sensitivity integration kit.Mycoplasma infection and drug susceptibility were analyzed.Results In the total of 1 9 530 suspected patients specimens,1 1 1 78 cases were positive with a positive rate 57.24%.The positive rate of ureaplasma urealyticum (Uu)and mycoplasma hominis (MH)was 44.63% and 0.44% respectively and the positive rate of Uu and Mh mixed infection was 12.1 7%.The positive rate of female was higher than that of male and the difference was statistically significant(P <0.05).The positive rate of mycoplasma in 2008-2014 was on the rise;The sensitive rate of mycoplasma to josamycin,doxycycline,minocycline element,clarithromycin was 88.57%,84.32%,76.09% and 71.53% respec-tively,mycoplasma was highly drug resistance to quinolone antibiotics;mixed infection resistance was higher than that of single in-fection;The number of drug resistance of Uu,MH and Uu+MH to 12 kinds of antibiotics increase.Conclusion Mycoplasma infec-tion in urogenital tract is mainly caused by Uu and Mh infection is in mixed infection way;josamycin,doxycycline is the first choice for treatment of mycoplasma in this region.Rational drug choise can be based on the drug susceptibility test.Multiple drug resist-ance of mycoplasma is serious and should be paid attention to.
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<p><b>OBJECTIVE</b>To obtain a specific antagonist of CXCR4, SDF-1P2G54 by mutating SDF-1 second proline (P) into glycin (G) and removing the α-helix of its C-terminal.</p><p><b>METHODS</b>SDF-1p2g54 gene amplified by PCR was inserted into the vector pET-30a (+) and transformed into Escherichia coli (E. coli) strain BL21. After IPTG induction of E. coli, the expressed recombinant protein was purified with nickel-affinity chromatography column under denaturing conditions and refolded with gradient dilution and ultra-filtration. The chemotactic effect of SDF-1P2G54 on Jurkat cells and its antagonistic effect against SDF-1 were determined by transwell assay; flow cytometry was used to assay the ability of SDF-1P2G54 to induce calcium influx and CXCR4 internalization in MOLT4 cells.</p><p><b>RESULTS</b>The recombinant protein SDF-1P2G54 completely lost the functions to activate CXCR4 or to induce transmembrane migration of Jurkat cells and calcium influx in MOLT4 cells, but maintained a high affinity to CXCR4. SDF-1P2G54 effectively inhibited the chemotactic effect of wild-type SDF-1 to Jurkat cells, and induced rapid CXCR4 internalization in MOLT4 cells.</p><p><b>CONCLUSION</b>SDF-1P2G54 is a new antagonist of CXCR4 with a potential value as an effective inhibitor of HIV-1 infection, cancer metastasis or other major diseases.</p>
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Humans , Cell Line , Chemokines, CXC , Genetics , Escherichia coli , Genetics , Metabolism , Mutant Proteins , Genetics , Receptors, CXCR4 , Recombinant Proteins , GeneticsABSTRACT
OBJECTIVE: To observe the promoting effects of blood-activating and stasis-removing Chinese drugs combined with vascular endothelial growth factor (VEGF) gene transfer on angiogenesis in ischemic necrosis of the femoral head. METHODS: Forty Japanese giant-ear rabbits were randomly divided into a control group, a model group, a Chinese drug group, a gene group, and a combined group. After 8 weeks of treatment, the rate of VEGF positive cell expression in the synovium of the femoral head was measured using the immunohistochemical method, and the number of blood vessels in the femoral head was measured by digital subtraction angiography. RESULTS: The rate of VEGF positive cell expression in the model group was significantly lower than that in the Chinese drug group (P < 0.05) and very significantly lower than those in other groups (P < 0.01); but in the combined group it was significantly higher than in the Chinese drug group (P < 0.05). The differences in the number of blood vessels in area A between the model group and other groups were not statistically significant. However, in area B, the number of blood vessels significantly increased in the control group, the gene group and the combined group as compared with the model group (P < 0.05), and in the combined group the number of blood vessels was significantly more than in the gene group (P < 0.05); but in the Chinese drug group it was not significantly different than the model group (P > 0.05). CONCLUSION: Either the blood-activating and stasis-removing Chinese drugs or VEGF gene transfer can promote the angiogenesis and building of collateral circulation for femoral head ischemic necrosis, and the combined therapy with Chinese drugs or VEGF gene transfer may show a better therapeutic effect. The present study provides an experimental basis for clinical application of the combined therapy with the blood-activating and stasis-removing Chinese drugs and VEGF gene transfer.
Subject(s)
Drugs, Chinese Herbal/therapeutic use , Femur Head Necrosis/therapy , Neovascularization, Physiologic , Vascular Endothelial Growth Factor A/metabolism , Angiography, Digital Subtraction , Animals , Combined Modality Therapy , Drugs, Chinese Herbal/administration & dosage , Femur Head/blood supply , Femur Head/metabolism , Femur Head/pathology , Femur Head Necrosis/etiology , Gene Transfer Techniques , Genetic Therapy/methods , Immunohistochemistry , Ischemia/complications , Rabbits , Random Allocation , Synovial Membrane/blood supply , Synovial Membrane/metabolism , Synovial Membrane/pathology , Vascular Endothelial Growth Factor A/geneticsABSTRACT
<p><b>BACKGROUND</b>SMAD proteins have recently been identified as the first family of putative transforming growth factor-beta1 (TGF-beta1) signal transducers. This study was to investigate the effects of TGF-beta1 and signal protein Smad3 on rat cardiac hypertrophy.</p><p><b>METHODS</b>The incorporation of [(3)H]-leucine was measured to determine the hypertrophy of cardiomyocyte incubated with different doses of TGF-beta1 in cultured neonatal cardiomyocytes. The model of rat cardiac hypertrophy was produced with constriction of the abdominal aorta. At different times after the operation, rats were killed, and their left ventricular mass index (LVMI) determined. The mRNA expression of TGF-beta1 and Smad3 of cultured cells and hypertrophic left ventricles were assessed by RT-PCR. The protein expression of Smad3 was assessed by Western blot.</p><p><b>RESULTS</b>In cultured neonatal cardiomyocytes, TGF-beta1 significantly promoted incorporation of [(3)H]-leucine. With the concentration of 3 pg/L, it increased the expression of Smad3 in mRNA and protein levels after 15 minutes, and continued for up to 8 hours of cultured cardiomyocytes. The LVMI and the expression of TGF-beta1 (mRNA) and Smad3 (mRNA and protein) of hypertrophic left ventricle were increased by day 3 after the operation and continued to the 4th week. The peak expression of these was in the second week after operation.</p><p><b>CONCLUSION</b>TGF-beta1 has positive effects on rat cardiomyocyte hypertrophy. Signal protein Smad3 could be related to the pathologic progression of rat cardiac hypertrophy.</p>
Subject(s)
Animals , Male , Rats , Aortic Coarctation , Metabolism , Cardiomegaly , Cells, Cultured , DNA-Binding Proteins , Physiology , Leucine , Metabolism , RNA, Messenger , Rats, Sprague-Dawley , Smad3 Protein , Trans-Activators , Physiology , Transforming Growth Factor beta , Genetics , Physiology , Transforming Growth Factor beta1ABSTRACT
Three forms of the major surface antigen (SAG1)of Toxoplasma gondii, that is the membrane form, the secrete form and the intracellular form, were constructed and used to immunize BALB/c mice. The humoral response in the mice immunized with the membrane form and the secrete form of SAG1 appeared earlier and stronger than those mice immunized with the intracellular form. Result from the challenging infection demonstrated that the protection in the mice immunized with the membrane and the secrete forms was also stronger than in the mice immunized with the intracellular form. We suggest that the immune efficiency of the three forms of SAG1 in the mouse model is different.
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Objective To identify the potential DNA vaccine candidate which can induce the protective immune response to Toxoplasma gondii by inoculating mice with plasmid DNAs encoding three different forms of P30 antigen (membranous secretory,and intracellular). Methods Three forms of recombinant plasmid: pcDNA3 P30Mb(contain the whole P30 gene sequence,including the gene encoding signal peptide and hydrophobic tail),pcDNA3 P30Se(contain the whole P30 gene sequence, without the gene encoding hydrophobic tail) and pcDNA3 P30In(contain the whole P30 gene sequence,without the gene encoding signal peptide) were constructed by PCR and subcloning technique. The mice were immunized with different forms of recombinant plasmids and IgG antibodies in the mice were detected by ELISA and Western blotting. Results Three forms of expression recombinant plasmid of Toxoplasma gondii P30 gene were successfully constructed. The P30 inserts were identified by restrictive enzyme digestion and sequencing. ELISA and Western blotting analysis demonstrated that specific IgG antibody could be induced in three immunized groups, but there was some difference in appearence time and intensity of IgG.Conclusion Genetically immunization of mice with the recombinant plasmids could elicit specific IgG antibodies. In respect to IgG response, the immune efficiency of the three forms of recombinant plasmids was different at the beginning (2 wk),but 4 wk later approximately same.