2.Changes in expression of calcium channel ?1B subunit mRNA in dorsal root ganglion neurons in a rat model of neuropathic pain
Chinese Journal of Anesthesiology 1994;0(03):-
Objective To determine the changes in expression of Ca2+ channel ?1B subunit mRNA in dorsal root ganglion (DRG) neurons in a rat model of neuropathic pain. Methods Twenty-four SD rats weighing 160-220g were randomly divided into 3 groups with 8 animals in each group: (1) neuropathic pain group; (2) sham-operated group and (3) control group. In neuropathic pain group the animals were anesthetized with intraperitoneal phenobarbital 50 mg?kg-1 . The left L5 spinal nerve was exposed and ligated and cut. In sham-operated the left L5 spinal nerve was exposed but not ligated and cut. Control group underwent no operation. Bilateral paw withdrawal threshold to von Frey hair stimulation was measured before and 7, 14 days after operation. The animals were killed on the 14th day after operation and the left L5 DRGs were obtained for determination of Ca2+ channel ?1B subunit mRNA by in situ hybridization. Results Mechanical allodynia developed in neuropathic pain group but not in sham-operated group. The expression of Ca2+ channel ?1B subunit mRNA was significantly lower in neuropathic pain group (0.033 ? 0.011) than in sham-operated group (0.065 ? 0.042) and control group (0.066 ? 0.034) (P
3.Substrate specificities of bile salt hydrolase 1 and its mutants from Lactobacillus salivarius.
Jie BI ; Fang FANG ; Yuying QIU ; Qingli YANG ; Jian CHEN
Chinese Journal of Biotechnology 2014;30(3):445-454
In order to analyze the correlation between critical residues in the catalytic centre of BSH and the enzyme substrate specificity, seven mutants of Lactobacillus salivarius bile salt hydrolase (BSH1) were constructed by using the Escherichia coli pET-20b(+) gene expression system, rational design and site-directed mutagenesis. These BSH1 mutants exhibited different hydrolytic activities against various conjugated bile salts through substrate specificities comparison. Among the residues being tested, Cys2 and Thr264 were deduced as key sites for BSH1 to catalyze taurocholic acid and glycocholic acid, respectively. Moreover, Cys2 and Thr264 were important for keeping the catalytic activity of BSH1. The high conservative Cys2 was not the only active site, other mutant amino acid sites were possibly involved in substrate binding. These mutant residues might influence the space and shape of the substrate-binding pockets or the channel size for substrate passing through and entering active site of BSH1, thus, the hydrolytic activity of BSH1 was changed to different conjugated bile salt.
Amidohydrolases
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genetics
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metabolism
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Bile Acids and Salts
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metabolism
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Escherichia coli
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metabolism
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Gene Expression
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Lactobacillus
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enzymology
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genetics
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Substrate Specificity
4.Changes of expression of FADD and Daxx following focal cerebral ischemia in rats
Yue-Qiang HU ; Bo XIAO ; Fang-Fang BI ; Ling DING ;
Chinese Journal of Neurology 2005;0(11):-
Objective To investigate the changes of expression of Fas-associated proteins named Fas-associated death domain protein(FADD)and death-associated protein(Daxx)in the ischemic penumbra following transient focal cerebra ischemia in rats.Methods ①Adult male Sprague-Dawley rats were randomly divided into the sham-operated group and the cerebral ischemia model group.Rats underwent right middle cerebral artery occlusion(MCAO)for 2 h and reperfusion for 1,3,6,12 and 24 h using an intraluminal suture technique.The expression of FADD and Daxx mRNA and protein were measured with methods of immunohistochemistry.Western blot and reverse transcription-polymerase chain reaction(RT- PCR)respectively were used in the ischemic penumbra of rats.②Double-label fluorescence confocal laser scanning microscopy(CLSM)was performed to monitor FADD and Daxx intracellular location before and after ischemia.Results RT-PCR,Immunohistochemistry,Western blot experiments indicated that a very low level of FADD mRNA and protein were detected in the cerebral cortex of sham rats.The expression level both of FADD mRNA and protein increased significantly at 3 h after reperfusion,peaked at 12 h,then declined markedly at 24 h in the ischemic penumbra of model rats.RT-PCR,Immunohistochemistry indicated that a relatively high level of Daxx mRNA was detected in the cerebral cotex of sham rats.The expression level of Daxx mRNA increased significantly at 3 h after reperfusion and persisted to 24 h at a high level,whose protein had a same change of expression level in the ischemic penumbra of model rats. Immunofluorescence double-staining laser scanning by CLSM showed that the immunoreactivity of FADD was located in cytoplasm,and the intracellular translocation of the immunoreactivity of Daxx from nucleus to cytoplasm was monitored by measuring the green fluorescence after ischemia.Conclusion The transient upregulation of FADD and the persistant high level of expression of Daxx may contribute to neuronal apoptosis following cerebral ischemia/reperfusion.
6.Diagnosis and treatment of primary splenic tumor: an analysis of 43 cases
Jin ZHANG ; Guoen FANG ; Jianwei BI
Chinese Journal of Hepatobiliary Surgery 2009;15(7):509-511
Objective To summarize the experience in diagnosis and treatment of primary splenic tumor. Methods The clinical data of 43 cases of primary splenic tumor patients treated in our hospital between January 1990 to January 2003 were retrospectively analyzed. Results The detectable rate of B-US was 95.3% and that of CT was 96.6%. All the 43 patients included 28 with benign tumors and 15 with malignant ones. The median follow-up period was 6.8 years(2 months-15 years). The 5-year overall survival of benign tumor was 100% and The 1-,3-and 5-year overall survival rates of malignant tumor were 80%, 53.3 % and 26.7%, respectively. Conclusion The image methods such as B-US and CT are the main diagnostic methods for primary splenic tumor. Early diagnosis, radical operation and combined therapy are important for improving the prognosis of the tumor.
7.Clinical observation on acupuncture at points of Shaoyang meridians plus moving cupping on the neck and shoulder for migraine
Journal of Acupuncture and Tuina Science 2017;15(5):377-381
Objective:To observe the efficacy of acupuncture at points of Shaoyang meridians plus moving cupping on neck and shoulder for migraine. Methods:A total of 64 migraine cases were randomly allocated into an observation group and a control group, 32 cases in each group. Random number table method was used in allocation. Acupuncture at points of Shaoyang meridians and cupping on neck and shoulder were used for cases in the observation group, which contain acupuncture 5 times a week and cupping once a week. Oral flunarizine hydrochloride capsules were used for cases in the control group, 10 mg for each dose, 1 dose a day. 2 weeks constitutes a course of treatment. The patients were treated for two courses of treatment in both groups. After that, the changes of visual analogue scale (VAS) and the migraine disability assessment questionnaire (MIDAS) were observed, as well as the clinical efficacy. Results:The total effective rate and recovery and marked effective rate in the observation group were 93.8% and 71.0% respectively, versus 78.1% and 43.8% in the control group, showing statistical significant differences (both P<0.05). There were significant decreases in VAS and MIDAS scores after treatments in both groups (both P<0.05). VAS and MIDAS scores in the observation group were significantly different from those in the control group (both P<0.05). Conclusion: Combining acupuncture at points of Shaoyang meridians and cupping on neck and shoulder can relieve headache and reduce influence of migraine on life. It can produce a better efficacy than oral flunarizine hydrochloride capsules in treating migraine patients.
8.Dynamic detection of C-reactive protein and procalcitonin to predict the occurrence of multiple organ dysfunction syndrome after multiple trauma
Jianguo WU ; Xuchao XUE ; Jianwei BI ; Guoen FANG
Chinese Journal of Postgraduates of Medicine 2010;33(35):1-3
Objective To investigate the relation of the peripheral blood C-reactive protein (CRP)and procalcitonin (PCT) and multiple organ dysfunction syndrome( MODS ) for patients with multiple trauma.Methods A total of 136 patients with multiple trauma were divided into MODS group (49 cases) and non MODS group (87 cases),and 50 healthy volunteers were chosen as control group. Peripheral blood CRP and PCT concentration were detected at different times. Results The highest concentration of peripheral blood CRP in MODS group was appeared on 48 h [(38.7 ± 2.7) mg/L], CRP concentrations in MODS group were significantly different with non MODS group and control group (P <0.05). The highest concentration of peripheral blood PCT in MODS group was appeared on 24 h [(20.3 ± 1.7)μ g/L], PCT concentrations in MODS group were significantly different with non MODS group and control group (P< 0.05). Conclusions CRP and PCT are relevant to the occurrence of MODS in trauma acute stage. The observation of peripheral blood CRP and PCT concentrations can predict the occurrence of MODS.
10.Change in calcium currents in isolated dorsal root ganglion neurons in rats with neuropathic pain
Fang LUO ; Hao-Sheng BI ; Lin RUAN ; Al ET
Chinese Journal of Anesthesiology 1994;0(06):-
Objective To examine the changes in calcium currents in isolated dorsal root ganglion (DRG) neurons in rats with neuropathic pain.Methods The neuropathic pain model was established by modified spinal nerve ligation (SNL) 2 to 4 weeks before electrophysiologic recording. The rat DRG neurons were enzymatically dissociated. Whole -cell patch clamp technique was used to record Ca2+ current.Results In large DRG neurons the mean peak value of electric current-voltage ( Ⅰ - Ⅴ) curve was decreased significantly from ( - 105?13) pA/ pF in control group ( n = 9) to ( - 66?10) pA/pF in neuropathic pain group ( n = 11) (P 0.05) . Conclusion In neuropathic rat Ca2+ currents in large DRG neurons are decreased and the voltage dependence of the fast component of inactivation is shifted to more depolarized potentials. These changes may contribute to hyperalgesia and allodynia of neuropathic pain.