1.Expression and significance of Maspin and IKKα in sinusoidal mucosa of rats with fungal rhinosinusitis
Chinese Journal of Comparative Medicine 2017;27(7):75-80,92
Objective To investigate the expression and significance of Maspin and IKKα in nasosinusoidal mucosa of rats with fungal rhinosinusitis (FRS).Methods A total of 40 SD rats were used to establish the FRS model, and randomly divided into nasal obstruction group, FRS group, immunosuppressive group and invasive FRS group, 10 rats in each group.Another 10 normal rats were used as control group.Mice in the control group were fed with normal diet.In the nasal obstruction group, the mice had only hemostatic cotton stuffed in the nasal cavity and injection of 0.9% NaCl in the abdominal and nasal cavities.In the FRS group, the mice were injected Aspergillus fumigatus spore suspension into the nasal cavity and 0.9% NaCl i.p.The mice of the immunosuppressive group were given cyclophosphamide i.p.and 0.9% NaCl injection into the nasal cavity.The invasive FRS group was injected with cyclophosphamide i.p.and Aspergillus fumigatus spore suspension into the nasal cavity.The serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA).The expression of Maspin and IKKα in nasosinusoidal mucosa was detected by immunohistochemical staining.The expression of Maspin mRNA and IKKα mRNA in the nasosinusoidal mucosa was detected by fluorescence quantitative PCR.Results The serum levels of IL-6 and TNF-α in different groups were significantly different (P< 0.05).The level of IL-6 in the FRS group was (69.3 ± 10.9) ng/L, significantly higher than those in the control group, nasal obstruction group, immunosuppressive group and invasive FRS group [(45.2 ± 7.1)ng/L, (46.4 ± 6.7) ng/L, (21.3 ± 4.5) ng/L, and (20.9 ± 4.3 ng/L)] (P < 0.05).The level of TNF-α in the FRS group was (30.4 ± 4.8) ng/L, significantly higher than those in the control group, nasal obstruction group, immunosuppressive group and invasive FRS group [(14.8 ± 2.7) ng/L, (13.9 ± 1.4) ng/L, (7.9 ± 0.6) ng/L, and (7.8 ± 0.4 ng/L)] (P < 0.05).The levels of IL-6 and TNF-α in the control group were significantly higher than those in the immunosuppressive group and invasive FRS group (P< 0.05).There was no significant difference between the immunosuppressive group and the invasive group (P> 0.05).Theresult of immunohistochemical staining showed that the protein expression of Maspin in the FRS group and invasive FRS group was significantly lower than that in the control group, nasal obstruction group and immunosuppressive group, while the expression of IKKα protein was significantly higher than that of control group, nasal obstruction group and immunosuppressive group (P< 0.05).The protein expression of Maspin in the invasive FRS group was significantly lower than that in the FRS group, by contrast, the expression of IKKα protein was significantly higher (P< 0.05).The PCRresult revealed that the expression levels of Maspin and IKKα mRNA were (0.217 ± 0.013) and (0.193 ± 0.012), significantly lower than that in the control, obstruction and immunosuppressive groups [(0.309 ± 0.021), (0.302 ± 0.017), and (0.293 ± 0.02)] (P< 0.05), while the expressions level of IKKα mRNA were significantly higher [(0.319 ± 0.043), (0.384 ± 0.048) vs (0.169 ± 0.015), (0.171 ± 0.018), and (0.175 ± 0.019)] (P< 0.05).Conclusions Down-regulation of Maspin expression after IKKα activation is the main cause of the onset of FRS, which may also be one of the mechanisms of invasive FRS.
2.Preliminary Analysis on TCM Syndrome Differentiation and Treatment about Diabetic Diarrhea
Qin LI ; Yanbin GAO ; Bingxin XIE
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(10):115-117
Diabetic diarrhea is one of the chronic complications of diabetes, which is particularly closely related to the spleen and kidney. Using these two organs as the theoretical foundation, the root of diabetic diarrhea can be grasped. Pathogenic dampness is responsible throughout the whole process of diabetic diarrhea. Using syndrome differentiation, deficiency is as the basic pattern and excess is the syndrome for diabetic diarrhea; diagnosis and treatment combine symptoms and syndromes, which not only particularly pay attention to protecting the spleen and kidney, but also clearing pathogenic dampness. At the same time, flexible medication is applied according to dynamic symptoms, which can achieve good efficacy.
3.The Analysis and the Strategy for the Influenced Factor of Doctorial Student's Resource
Yue-Wen QIN ; Wei LI ; Li-Li DAI ; Peng XIE ;
Chinese Journal of Medical Education Research 2006;0(10):-
While the number of the enrolled postgraduate students is continuously increased and the various reforms go in depth, there appear some new problems and contradictions in the education of doctorial students.Among those problems,the lack of the doctorial student resource,especially the lack of the excellent doctorial student resource,is the most prominent problem that re- stricts the progress of the doctorial student education.In order to solve these problems,this article analyzes the factors influencing the resource of the students and put forward the strategy to deal with these factors.
4.Effects of propofol anesthesia on long-term cognitive function in neonatal rats
Li LI ; Chunjie LIAO ; Yi QIN ; Yubo XIE
Chinese Journal of Anesthesiology 2012;(10):1204-1207
Objective To investigate the effects of different doses of propofol anesthesia on the long-term cognitive function in neonatal rats.Methods One hundred 7-day-old male Sprague-Dawley rats,weighing 9-18 g,were randomly divided into 5 groups (n =20 each) ∶ control group (C group) and propofol 25,50,100 and 200mg/kg groups (groups P1-4,respectively).Groups P1 and P2 received intraperitoneal propofol 25 and 50 mg/kg,respectively.Groups P3 and P4 received intraperitoneal propofol 100 and 200 mg/kg,respectively,and after righting reflex completely recovered,an increment of propofol 50 mg/kg was given until the total amount was finished.Five animals in each were chosen and arterial blood samples were obtained immediately after the animals were fully awake for blood gas analysis.The rats were then continuously fed.When the rats were 9 weeks old,the spatial learning and memory function was tested by Morris water maze.The animals were then sacrificed and their brains were removed for detection of the expression of nerve growth factor (NGF) and Caspase-9 protein and mRNA in hippocampal tissues (by Western blot analysis and RT-PCR) and for microscopic examination of the ultrastructure of hippocampal neurons.Results There was no significant difference in blood gas analysis index between the 5 groups (P > 0.05).Compared with group C,no significant change was found in the escape latency and frequency of crossing the original platform in group P1,the escape latency was prolonged and the frequency of crossing the original platform was decreased in groups P2-4,the expression of NGF protein and mRNA was down-regulated and the expression of Caspase-9 protein and mRNA was up-regulated in groups P1-4 (P < 0.05).The escape latency was gradually prolonged,the expression of NGF protein and mRNA was gradually down-regulated and the expression of Caspase-9 protein and mRNA was gradually up-regulated in groups P1-4.The frequency of crossing the original platform was decreased in groups P2-4 compared with group P1 (P < 0.05).There was no significant difference in the frequency of crossing the original platform between groups P2-4 (P > 0.05).Nucleus condensation,chromatin condensation,nuclear fragmentation and apoptotic bodies were observed in groups P2-4.Conclusion Propofol anesthesia can impair the long-term cognitive function in neonatal rats,the effect is related to the dose,and inhibition of NGF expression and increase in the activity of Caspase-9 may be involved in the mechanism.
5.Effect of fentanyl on viability of human gastric carcinoma cell line MGC-803
Yi QIN ; Li LI ; Jing CHEN ; Yubo XIE
Chinese Journal of Anesthesiology 2010;30(6):705-707
Objective To investigate the effect of fentanyl on the viability of human gastric cancer cell line MGC-803. Methods The human gastric cancer cell line MGC-803 was purchased from Cell Biology Research Institute, Chinese Academy of Sciences, and cultured in DMEM liquid culture medium. The cells were seeded in 6-well or 96-well plates and divided into 3 groups (n = 60 wells each): group Ⅰ normal control (group C); group Ⅱ and Ⅲ were exposed to fentanyl 0.01 and 1.00 μmol/L respectively (group F1, F2). The viability of the cells was detected by MTT assay after being incubated with fentanyl for 12, 24, 36, 48, 60 and 72 h. The cell cycle progression and apoptosis were assessed by flow cytometry and the ulrastructure of the cells was examined with transmission electron microscope after being incubated with fentanyl for 24 h. The proliferation of the cells was determined by colony formation assay at 7 day of incubation with fentanyl. Results The viability and proliferation of the cells and the proportion of the cells in S phase were significantly lower, while the proportion of the cella in G2/M phase and the apoptotic rate were higher in group F1 and F2 than in group C but no significant difference was found between group F1 and F2. The nuclear evelope was intact, the nucleolus and chromosomes were clearly visible in group C, while in group F1 and F2 fregmentation of nuclear envelope and nucleolus, chromatin condensation and apoptotic bodies were observed in group F2. Conclusion Fentanyl can inhibit the viability of human gastric cancer cells by its pro-apoptosis inducing effect.
6.Effect of morphine on growth of subcutaneous tumor of human gastric cancer cells in nude mice
Lifang ZHOU ; Yi QIN ; Li LI ; Yilan YANG ; Yubo XIE
Chinese Journal of Anesthesiology 2016;36(5):578-580
Objective To evaluate the effect of morphine on the growth of subcutaneous tumor of human gastric cancer cells in nude mice.Methods Thirty SPF male BALB/C nude mice,aged 4-5 weeks,weighing 15-20 g,in which the model of subcutaneous tumor of human gastric cancer cell line MGC-803 was established,were randomly divided into 3 groups (n=10 each) using a random number table:control group (group C),normal saline group (group N),and morphine group (group M).The mice in group C received no treatment.Morphine 20 mg/kg was intraperitoneally injected once a day for 14 consecutive days in group M,while normal saline 1.5 ml/kg was given instead of morphine in group N.The caliper was used to measure the tumor size every 2 days starting from 3 days after beginning of administration,and the relative tumor volume was calculated.The nude mice were sacrificed on 15th day,and the tumor tissues were obtained for determination of nuclear factor-kappa B activity and Bcl-2 and Bax protein and mRNA expression by real-time reverse transcriptase polymerase chain reaction and Western blot.Results Compared with group C,the relative tumor volume was significantly decreased,the activity of nuclear factor-kappa B in tumor tissues was significantly decreased,the expression of Bcl-2 protein and mRNA was significantly down-regulated,and the expression of Bax protein and mRNA was significantly up-regulated at each time point in group M (P<0.05),and no significant change was found in each parameter mentioned above in group N (P>0.05).Conclusion Morphine can inhibit the growth of subcutaneous tumor of human gastric cancer cells in nude mice,and the mechanism is associated with promotion of apoptosis in tumor cells.
7.Effect of naloxone in combination with morphine on the growth of subcutaneous tumor of human gastric cancer MGC-803 cells in nude mice
Hao WANG ; Yi QIN ; Yubo XIE ; Li LI
The Journal of Clinical Anesthesiology 2017;33(3):277-280
Objective To observe the effects of naloxone in combination with morphine on the growth of subcutaneous tumor of human gastric cancer MGC-803 cells in nude mice.Methods The model of subcutaneous tumor of human gastric cancer MGC-803 cells in nude mice was established.Fifty nude mice were randomly divided into 5 groups: control group (group C),normal saline group (group S),20 mg/kg morphine group (group M),and 1 mg/kg naloxone group (group N),and 1 mg/kg naloxone+20 mg/kg morphine group (group NM).The mice in group C received no treatment,while the mice in group S,group M,group N and group NM were injected with 1.5 ml/kg saline,20 mg/kg morphine,1 mg/kg naloxone,and 1 mg/kg naloxone+20 mg/kg morphine per day,respectively.The caliper was used to measure the tumor sizes every the other day.The mice in each group received intraperitoneal injection of the drugs for 2 week.Then the relative volume (RTV) of tumor was calculated.The expression of Cyclin D1,VEGF,MMP-9 mRNA and proteins were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR),immunochemistry staining and Western blot.Results RTV in group M (2.21±0.62)% was significantly lower than that in group C (3.16±0.68)%,group S (2.98±0.61)%,group N (3.16±0.35)% and group NM (2.64±0.37)% (P<0.05).RTV in group NM was significantly lower than that in group C,group S and group N (P<0.05).Compared with group C,the expression of Cyclin D1,VEGF,and MMP-9 in group M were significantly decreased (P<0.05).The organizational structure of the subcutaneous tumor in groups C,S,N and NM was almost normal.Cytoplasm vacuolization,disruption of nuclear membrane and chromatin margination were occured in group M.While the level of Cyclin D1,VEGF,and MMP-9 in group NM was increased compared to group M (P<0.05).Conclusion Morphine could inhibit the growth of subcutaneous tumor of human gastric cancer MGC-803 cells in nude mice by downregulating the expression of Cyclin D1,VEGF,and MMP-9.Naloxone could antagonize the anti-growth effects of morphine.
8.Effects of electroacupuncture at Chize (LU 5) versus Shangjuxu (ST 37) in rats with ulcerative colitis
Qin LUO ; Zhitong LI ; Weining YANG ; Shengjie LI ; Jieping XIE
Journal of Acupuncture and Tuina Science 2015;(4):207-211
Objective:To compare the effects between electroacupuncture (EA) at Chize (LU 5, the He-Sea point of the Lung Meridian) and Shangjuxu (ST 37, the lower He-Sea point of the large intestine) in rats with ulcerative colitis (UC) on the variations of mesenteric microcirculation and vasoactive intestinal peptide (VIP) in the colon, lung, and hypothalamus. The relative specificity of acupoints was also explored.
Methods: A total of 28 male Wistar rats were randomized into a normal group, a model group, a Chize (LU 5) group and a Shangjuxu (ST 37) group, 7 rats in each group. The UC model was established by enema with acetic acid. Since the third day after modeling, rats in the Chize (LU 5) group and Shangjuxu (ST 37) group respectively received EA at Chize (LU 5) and Shangjuxu (ST 37), 15 min each time for successive 7 d. The variations of mesenteric microvascular calibers and blood flow status were observed by a microcirculation microscopic tester; VIP in the colon, lung and hypothalamus was measured by radioimmunoassay.
Results:Compared with the normal group, the mesenteric microvascular calibers were significantly expanded in the model group (P<0.05); there was no significant difference between the model group and Chize (LU 5) group (P>0.05); compared with the model group and Chize (LU 5) group, the calibers were obviously shrunk in Shangjuxu (ST 37) group (P<0.05). The four groups showed no significant inter-group differences in comparing blood flow status (P>0.05). The colonic VIP levels in the model group and Chize (LU 5) group were significantly higher than that in the normal group (P<0.01,P<0.05); the VIP level in Shangjuxu (ST 37) group was markedly lower than that in the model group (P<0.01). There were no significant differences among the four groups in comparing VIP level in lung and hypothalamus (P>0.05).
Conclusion:The effects of Chize (LU 5) and Shangjuxu (ST 37) were different in treating UC. Shangjuxu (ST 37) showed a more significant efficacy in down-regulating VIP in the colon and regulating mesenteric microcirculation, while the effects of Chize (LU 5) were not obvious.
9.Two-step cluster analysis and corresponding analysis in the syndrome type of knee osteoarthritis
Bin HU ; Xingwen XIE ; Ning LI ; Jin HUANG ; Linyuan QIN
Chinese Journal of Tissue Engineering Research 2014;(11):1799-1804
BACKGROUND:Both correspondence analysis and two-step cluster analysis are high-grade statistical analysis, the introduction of these analyses into the research on traditional Chinese medicine (TCM) syndrome type of knee osteoarthritis wil provide objective evidence for the standardization and normalization of TCM syndrome type, through the combination of mathematical statistical principle and TCM syndrome type.
OBJECTIVE:To explore distribution characteristics of knee osteoarthritis TCM syndrome type using correspondence analysis and two-step cluster analysis.
METHODS:The clinical symptoms of 200 patients with knees osteoarthritis were investigated through a knee osteoarthritis symptoms questionnaire. According to the criteria for three kinds of syndrome type issued in Diagnostic Criteria for TCM Syndrome, the characteristics of each syndrome were analyzed using two-step cluster analysis and corresponding analysis. Then knee osteoarthritis TCM syndrome type characteristics were defined.
RESULTS AND CONCLUSION:Cluster analysis is ineffective for the syndrome type, which is not present in the Diagnostic Criteria for TCM Syndrome. Corresponding analysis showed that, in addition to kidney marrow deficiency syndrome (50.5%), yang deficiency and congealing syndrome (13.5%), and blood stasis syndrome (23%), concurrent syndromes were also found, including kidney marrow deficiency combined yang deficiency and congealing syndrome (6.5%), yang deficiency and congealing combined blood stasis syndrome (3%), kidney marrow deficiency combined blood stasis syndrome (3.5%). Therefore we performed corresponding analysis. After analyzing the syndromes at 0.5, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5 radius, the most reasonable syndrome was those at 1.1 radius by corresponding analysis. Corresponding analysis is a scientific method for the classification of knee osteoarthritis syndrome.
10.Expressions of calpain 1 and caspase-3 in lichen planus lesions and their relationship with apoptosis in keratinocytes
Xin XIE ; Xiyu GAI ; Xiaojuan QIN ; Lin ZHANG ; Li CHAI
Chinese Journal of Dermatology 2012;(12):859-861
Objective To quantify the expression levels of calpain 1 and caspase-3 in lichen planus (LP) lesions and their relationship with the apoptosis in keratinocytes.Methods Biopsy samples were obtained from the lesions of 20 patients with LP and normal skin of 10 healthy controls,and embedded in paraffin.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) was used to evaluate the apoptosis in keratinocytes,and immunohistochemical staining to detect the expressions of calpain 1 and caspase-3,in these tissue specimens.Data were processed by SPSS 13.0 software.Comparison analysis was carried out by t test for apoptosis index,and by rank sum test for the expressions of calpain 1 and caspase-3.Spearman correlation analysis was conducted to evaluate the relationship between these parameters.Results The apoptosis index of keratinocytes was higher in LP lesions than in the normal skin (67.59 ± 13.50 vs.28.26 ± 7.56,t =8.52,P < 0.01).Significantly increased expressions of calpain 1 and caspase-3 were observed in the epidermis of LP lesions compared with the normal skin (T =78.00 and 77.00,respectively,both P < 0.01).The expressions of both calpain 1 and caspase-3 were positively correlated with apoptosis index of keratinocytes (r =0.71 and 0.74,respectively,both P < 0.01).Conclusions The expressions of calpain 1 and caspase-3 are upregulated in LP lesions,which may be closely associated with the accelerated apoptosis in keratinocytes.