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AIM: To observe the effects of Tangshen formula (TSF) treatment on lipid efflux and uptake in sodium palmitate (PA) induced RAW264.7 macrophages. METHODS: After 200 μmol/L PA induced RAW264.7 macrophages, TSF and PGC-1α-siRNA were given to intervene respectively. The lipid content in the cells was detected by ELISA kit; intracellular lipid droplet deposition was detected by BODIPY 493/503 and Filipin staining. Western blot and Real-time PCR were used to detect the expression of PGC-1α, LXR, ABCA1 and CD36. RESULTS: TSF diminished the levels of TC, TG and intracellular lipid droplet deposition in PA-induced RAW264.7 macrophages. Western blot and Real-time PCR analysis showed that TSF could up-regulate the expression of PGC-1α, LXR, ABCA1 and down-regulate the expression of CD36. Furthermore, silencing PCG-1α by SiRNA significantly suppressed the effects of upregulating the expression of PGC-1α, LXR and ABCA1, and downregulating the CD36 expression with TSF treatment. CONCLUSION: TSF may extenuate intracellular lipid droplet deposition in macrophages by upregulating cholesterol efflux through activating the PGC-1α/LXR/ABCA1 pathway and inhibiting lipid uptake through down-regulateing the expression of CD36.
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Objective:To evaluate the correlation of inter-individual variation of cyclosporine dosage and blood concentration and CYP3A4 and CYP3A5 polymorphism in renal transplant recipients. Methods:Two hundred and twenty-one renal transplant recipients treated with cyclosporine were genotyped for CYP3A4 rs4646437C>T and CYP3A5 6986G>A using ligase detection reactions. The effects of genetic polymorphisms of CYP3A4 and CYP3A5 on cyclosporine trough concentration (C0/D) and 2 h post-dose concentra-tion (C2/D) during the period of 6 months, 6-24 months and above 24 months after renal transplant were studied. Results:CYP3A5 6986G>A genotype affected C0/D during the period of 6 months, 6-24 months and beyond 24 months (PGA>AA. CYP3A4 rs4646437C>T and CYP3A5 6986G>A genotype affected C2/D during the period above 24 months (PCT>TT and CYP3A5 6986GG>GA>AA. Conclusion: CYP3A5 6986G>A genotype affects C0/D and C2/D of cyclosporine, and CYP3A4 rs4646437 C>T genotype affects C2/D of cyclosporine. The effects of genotypes are varied in different stages.
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Objective To evaluate and compare C5 palsy and closure of the opened lamina after expansive open-door Laminoplasty (EOLP) with miniplate or suture/anchor fixation.Methods Between January 2011 and January 2013,a total of 142 patients with cervical myelopathy who were treated by EOLP were divided into hinge-side fixation group (fixed with suture/anchor,78 cases)and open-side fixation group (fixed with miniplate,64 cases).The Japanese Orthopaedic Association (JOA) score was used for neurological assessment and recovery rate (RR) counting.Opening angles,cervical curvature index (CCI),posterior shifting of spinal cord (PSSC) and severity of cord compression were recorded and compared.Results All patients in both group were followed up for more than 12 months.All incisions healed by first intention.C5 palsy occurred in 9 patients (9/78,11.5％) of hingeside fixation group,and 1 patients (1/64,1.6％) of open-side fixation group,showing significant difference (P=0.047).Opening angles and PSSC in hinge-side fixation group were greater than that in open-side fixation group.PSSC of 10 patients with C5 palsy were 3.97±1.19 mm,and greater than that of other patients without C5 palsy 2.57± 1.01 mm.There was no significant difference in CCI before (12.23％±3.70％,11.38％±4.29％) and 1 week (12.12％±3.77％,11.31％±4.35％) after operation.No significant difference was found in JOA scores (12.35±1.09,13.55±0.91),JOA improvement rate (64.24％±9.49％,61.78％±11.48％) and cord compression (0.74±0.71,0.75±0.67) at 12 months after operation.In 6 months postoperatively,27％ of patients in hinge-side fixation group,none in open-side fixation group were identified with 10％ decrease or more in opening angles of lamina.Conclusion EOLP with miniplate fixation has the same clinical outcome as fixed with suture/anchor,but will reduce the incidence of C5 palsy and prevent further closure of the opened lamina.
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BACKGROUND:To in vitro isolate neural stem cel s with high purity and uniform biological properties and to establish a complete set of neural stem cel culture system is the basis for neural stem cel research. OBJECTIVE:To establish an isolation and culture system for neural stem cel s from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cel s. METHODS:Neural stem cel s were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cel s. 10%fetal bovine serum was used to induce differentiation of neural stem cel s. Neural stem cel s and their differentiated products were identified by immunofluorescent staining of Nestin, CD133,β-TubulinIII, glial fibril ary acidic protein. RESULTS AND CONCLUSION:The neural stem cel obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cel could differentiation toβ-tubulinIII or glial fibril ary acidic protein positive cel s that were neurons and astrocytes. This experiment has successful y established the neural stem cel isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cel s.
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AIM To establish a dysmenorrhea model in mice. METHODS The mice were given with some kinds of oestrogens once a day for 3～25 days. On the last day, the mice were injected intraperitoneally with oxytocin and the number of twisting body was recorded to evaluate the intensity of dysmenorrhea. The optimum conditions to establish the model was analysed statisticly. RESULTS The optimum oestrogens was stilbestrol. Stibestrol should be given for 12～15 days. The regression equation of the dose effect curve of stibestrol was =0 03?0 04 X, r =0 9688. The optimum dosage of oxytocin was 20 U?kg -1 . The dysmenorrhea model in mice could be preserved for about 7 days. 90% of the twisting body reactions concentrated in 0～30 minutes after oxytocin was given. The effect of oxytocin (20 U?kg -1 ) had significent difference with that of prostaoglantin (1 3 mg?kg -1 ). The test of uterus in vivo showed that stilbestrol could increase the uterine contraction frequency and strengthen the contractility. The dysmenorrhea model in mice was testified by some anti dysmenorrhea drugs. CONCLUSION Compared with the hypodermic implantation in rats, the dysmenorrhea model in mice was simple, reliable, economical and testifiable,etc.
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Objective To investigate the influence of case environment, extracting position and method on STR typing of nail DNA. Method Decayed body nails were selected from cases. Nails nuclear DNA were extracted from different positions, digestion system and time. After phenol/chloroform extracting and PCR reaction, the STR typing had been studied. Results Nail samples from middle and root positions offer satisfactory results. Conclusion The nail is a useful sample for forensic cases.