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Objective To optimize the extraction technology of total triterpenoid from root of Rose odorata var.gigantean (TTROG) by orthogonal test combined with the contraction effect of TTROG on the isolated intestinal smooth muscle of rats in vitro.Methods UV spectrophotometric method was used to determine the contents of total triterpenoids in the TTROG extractive at the wavelength of 550 nm by taking ursolic acid as standard substance,and vanillin acetic acid as chromogenic reagent.The extraction rate of total triterpenoids was used as index to evaluate the technology based on single factor test,in which three factors were considered as follows:the concentration of extraction solvent,ratio of material to liquid,extraction time,and their interaction on extraction were studied by orthogonal experimental design.The inhibition effect of different extracts obtained from the optimized extraction process on the contraction of intestinal smooth muscle were recorded by tension transducer to the BL-420 biological experimental multi-channel physiological signal acquisition and processing system.The extraction process of TTROG was evaluated by the combination of biological activity and extraction rate with weighting method.Results The optimal extraction conditions of TTROG were as follows:extraction solvent 80％ ethanol,solid-liquid ratio 1∶10,extraction time for 2 h,three times and extraction temperature of 80 ℃.The optimized extraction rate could reach 42.12 mg/g.TTROG obtained using the optimized method showed significantly contraction effect on rat intestinal smooth muscle with dose effect dependence,and the effect on jejunum was the strongest,and the inhibition rate was 41.96％.Conclusion The optimized extraction technology is stable and effective with high extraction rate.TTROG showed the significant inhibitory function on contraction of isolated rat intestinal smooth muscle.
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Objective Type 1 diabetes mice model was established to investigate the changes of key enzymes involved in testosterone synthesis in testes of early diabetic mice.Methods Tatolly 20 male C57 mice were randomly divided into two groups:control and diabetic groups,and the diabetes mice were ip administered with a single dose of 150 mg/kg Streptozotocin.Four weeks after confirmation of diabetic model,the serum and testis were collected for further study.The qRT-PCR method was used to measure the expression of LHR and steroidogenesis synthetase StAR,P450scc,3β-HSD6,P450c17a1,and 17β-HSD3 mRNA.ELISA assay was performed to measure the levels of testosterone and luteinizing hormone (LH) in serum,and the enzymatic activities of 3β-HSD1,1P450c17 and 17β-HSD3 in testis tissue.Results Compared to control group,the levels of testosterone and LH of diabetic group declined significantly (P ＜ 0.05) after four weeks.The mRNA levels of LHR,StAR,CYP11a1,Hsd3b6,CYP17a1 and Hsd17b3,and enzymatic activities of 3β-HSD6,P450c17 and 17β-HSD3 were also decreased significantly compared with control group (P ＜ 0.05,0.01 and 0.001).Conclusion Expression of key enzymes of testosterone synthesis in testis of early diabetic mice decreases significantly.
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Objective To investigate whether pro-inflammatory cytokines ( PICs) in the paraventricular nucleus ( PVN) regulate the enhanced sympathetic activities in spontaneously hypertensive rats ( SHR) , and whether N-methyl-Daspartate receptor ( NMDAR ) in PVN mediate the effects of PICs on sympathetic activities. Methods SHR and normotensive wistar-Kyoto( WKY) rats were used in this experiment. TNF receptor and IL-1β receptor ( IL-1RI) protein levels were measured by Western blot. PICs, including TNF-α and IL-1β levels were measured by ELISA. Rats were placed in a stereotaxic instrument to complete the microinjection of drugs. The coordinates for the PVN were determined according to the Paxinos and Watson rat atlas. The raw RSNA and integrated RSNA were simultaneously recorded on a PowerLab data acquisition system. The right carotid artery was cannulated for recording of mean arterial pressure ( MAP) . Results TNF-α receptor p55TNFR, p75TNFR and IL-1βreceptor IL-1RI protein expression and TNF-αand IL-1βlevels in PVN were all increased in SHR compared with WKY rats (P< 0. 05). Bilateral microinjection of etanercept or IL-1ra into PVN to block the effects of TNF-αor IL-1βdecreased the sympathetic activities in SHR rats significantly (P< 0. 05). Bilateral microinjection of NMDAR blockers, both DL-2amino-5-phosphonovaleri acid ( APV) and MK-801 ( Dizocilpine) into PVN decreased the RSNA and MAP in both SHR and WKY rats. APV or MK 801 caused greater decreases in RSNA and MAP in SHR than WKY rats. In addition, pretreatment with APV or MK 801 attenuated the increased RSNA and MAP caused by microinjection of TNF-αor IL-1βinto PVN to a lower level in SHR than in WKY rats (P< 0. 05). Conclusions TNF and IL-1βreceptor protein as well as TNF-αand IL-1βcytokines levels in PVN are all increased in SHR rats. NMDAR in PVN mediates enhanced sympathetic activities and elevated blood pressure caused by TNF-αand IL-1βin SHR.
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A device to produce low temperature plasma ( LTP) was designed and constructed to serve as the ion source of a high resolution mass spectrometry, and was applied to qualitatively analyze the steroid samples. In comparison with conventional electrospray ionization mass spectrometry, low temperature plasma mass spectrometry ( LTP-MS) had some advantages such as simple sample pretreatment and less interference. Mass spectrometry and tandem mass spectrometry were used to characterize the steroid samples in this research, and it was found that the structural stability of each steroid sample was presented in its mass spectrum, while in the tandem mass spectra there were more fragments of H2 O lost. And then the fragmentation process of typical steroid samples in collision induced dissociation ( CID ) was discussed based on theoretical calculation. In addition, by comparing tandem mass spectrometry and the fragmentation process, a pair of isomers of testosterone and dehydroepiandrosterone could be distinguished successfully.
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·AIMS:To investigate the etiological factors and various effects of severe vitreous hemorrhage ( VH ) in Northern China.·METHODS:Files on patients presenting with VH treated with vitrectomy between January 2011 and January 2014 were retrieved from medical records.·RESULTS:A total of 1335 eyes of 1275 patients ( 735 males, 540 females) presenting with VH were included in this study. Proliferative diabetic retinopathy ( PDR ) , retinal vein occlusion ( RVO) , either retinal detachment or retinal hole (RD/RH), ocular trauma, Eales disease, and either age- related macular degeneration or polypoidal choroidal vasculopathy ( AMD/PCV ) constituted the etiology of VH in more than 90% of the patients. The most common causes of VH were ocular trauma ( 40%) , PDR (19. 5%), and Eales disease (19. 1%) in the youth group, PDR (34. 4%), RVO (30. 8%), and RD/RH (12. 2%) in the middle-aged group, and RVO ( 35. 7%) , PDR ( 26. 6%) , RD/RH (14. 6%), and AMD/PCV (8%) in the elder group.· CONCLUSION: PDR, RVO, and ocular trauma are usually the main causes of VH. Within each group, the most common causes of VH were ocular trauma and Eales disease in the youth group, PDR, RVO, and RD/RH in the middle-aged group, and RVO, PDR, RD/RH, and AMD/PCV in the elder group. In addition, we found that males with ocular trauma are at high risk for VH, PDR and Eales disease often present bilateral VHs, and PDR and RVO have a high risk of recurrence.
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Secondary lymphoid-tissue chemokine (SLC) is a type of CC chemokine identified by searching the Expressed Sequence Tag (EST) database. The full-length SLC gene was synthesized based on human SLC sequence using SOE-PCR. The sequenced SLC gene was cloned into expression vector pTMF and pALM, which used to transform Escherichia coli. Then the E. coli was cultured and induced according to protocol. The expressed target protein was identified by Western blotting. The target protein was expressed as soluble protein as well as inclusion bodies, the ratio of these two forms target protein varied with the difference conditions of culture and induction. The target protein was purified with the methods of nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography. The results of electrophoresis of the purified target protein showed that the molecular weight was larger than the predicted molecular weight.