1.Oxidative stress and interleukin-1β in acute gouty arthritis and the efficacy of probucol treatment
Ying YUAN ; Yan LUO ; Changgui LI
Chinese Journal of Endocrinology and Metabolism 2010;26(3):225-228
Objective To investigate the role of malondialdehyde(MDA),superoxide dismutase(SOD),and interleukin-1β (IL-1β) played in the pathogenesis of acute gouty arthritis and the treatment efficacy of probucol.Methods Acute gouty arthritis model in rat was established with injection of monosodium urate monohydrate (MSU) into the ankle joint cavity,then the joint swelling index was observed periodically,and MDA,SOD,IL-1β,and white blood cell count in the synovial fluid were determined by 72 h.Results The joint swelling index gradually increased,reaching a peak by 11h,and this finding was most obvious in the model group.The joint swelling index in the group treated with high-dose of probucol was significantly lower than that in the model group by 48 h(P<0.05).The joint swelling indices in the groups treated with moderate and high doses of probucol were lower than that in the model group by 72 h(P<0.05).The values of MDA in the moderate and high-dose groups were (14.45±3.11) and (11.54 ±3.10) nmol/ml respectively,being significantly lower than that in the model group[(24.46±4.27) nmol/ml,P<0.01].The amounts of SOD in the moderate and high-dose groups were (25.56±4.51) and (44.61±4.11)U/ml respectively,being higher than that in the model group[(13.32±2.02) U/ml,P<0.01].The amount of IL-1β in the high-dose group was lower than that in the model group[(15.41 ± 3.12 vs 19.87±3.99)pg,P<0.05].The white blood cell counts in the moderate and high-dose groups were (11.08±1.64)×l0~7/L and (7.43±1.52) x 10~7/L,being lower than that in the model group[(14.18±2.30)×10~7/L,P<0.01].Conclusion The levels of MDA and IL-1β in the synovial fluid were raised,while SOD was significantly decreased in acute gouty arthritis.Moderate and high-doses of probucol can effectively control acute gouty arthritis attack.The efficacy of high-dose probucol was equal to that of colchicines.
3.Principles and progress in treatment of talus fracture
Chinese Journal of Orthopaedic Trauma 2021;23(4):329-336
Although talus fractures are not common among foot and ankle injuries, their treatment is difficult because of their complications and poor prognosis. They can be treated conservatively or surgically, with a variety of treatment protocols. At present, the main strategy of surgical treatment is to use strong and effective fixation to restore anatomic structure of the talus and preserve blood supply to the talus as much as possible so that deformity can be avoided and early healing be facilitated. This paper reviews the difficulties and current situation, and summarizes principles and the latest progress in clinical treatment of talus fractures, hoping to provide useful references for the treatment.
4.Influence of high-mobility group box 1 on proliferation of neural stem cell in pre-infarction cortex of focal cerebral ischemia/reperfusion model rats
Man LI ; Yong LUO ; Yuan LI ; Lin SUN
Chinese Journal of Pathophysiology 2014;(9):1616-1621
AIM:To investigate the influence of high-mobility group box 1 (HMGB1) on the proliferation of neural stem cells in peri-infarction cortex of focal cerebral ischemia/reperfusion model rats .METHODS: Male SD rats (n=48) were randomly divided into sham group , ischemia/reperfusion (I/R) group, RNA interference group and nega-tive interference group .The rat middle cerebral artery was blocked to establish focal cerebral I /R model ( ischemia for 1 h and reperfusion for 7 d).Lentivirus vector of HMGB1 shRNA was used to suppress the HMGB1 protein expression in the rat brain.The effect of RNA interference was evaluated by the methods of double-immunofluorescence labeling of HMGB 1/GFAP and Western blotting .The proliferation of neural stem cells in the peri-infarction cortex was assessed by double labe-ling of BrdU/nestin.RESULTS: The protein expression of HMGB1 in I/R group was much higher than those in sham group (P<0.05).RNA interference effectively inhibited the HMGB1 expression (P<0.05).Double labeled BrdU/nestin positive cells in I/R group were more than that in sham group (P<0.05).The double labeled BrdU/nestin positive cells were significantly decreased in RNA interference group (P<0.05).CONCLUSION:Focal cerebral ischemia/reperfusion injury promotes the proliferation of neural stem cells in peri-infarction cortex by increasing HMGB 1 protein level .
5.Effect of antidepressants on the hippocampal neurogenesis in chronically stressed mice
Yunfeng LI ; Yanqin LIU ; Youzhi ZHANG ; Li YUAN ; Zhipu LUO ;
Chinese Pharmacological Bulletin 1986;0(04):-
AIM To explore possible action mechanism of antidepressants. METHODS Using flow cytometry, the cell proliferation was detected. The proliferation of hippocampal progenitor cells and level of brain derived neurotrophic factor (BDNF) were measured by immunohistochemistry. RESULTS Treatment with N methyl D aspartate (NMDA) 600 ?mol?L -1 for 3 d significantly decreased the percentage of S phase in PC12 cells, while in the presence of classical antidepressants, desipramine (DIM) or fluoxetine (FLU) 1, 5 ?mol?L -1 , the percentage of S phase increased. Furthermore, the proliferation of hippocampal progenitor cells, as well as the BDNF level in dentate gyrus (subgranular zone) significantly decreased in chronically stressed mice for 24 d, while chronic administration with DIM or FLU 10 mg?kg -1 (ip) normalized it. Meanwhile, the BDNF level in dentate gyrus also elevated after DIM or FLU treatments. CONCLUSION Up regulation of the hippocampal neurogenesis is the common action mechanism for antidepressants, which may be closely related to the elevation of BDNF level at the same time.
6.Correlation between the expression of gastric cancer tumor markers in preoperative serum and the risk of radical resection
Xiyu YUAN ; Qingxian LI ; Ping LUO ; Lihua LI
Clinical Medicine of China 2021;37(1):67-73
Objective:To study the relationship between the expression of tumor markers and radical resection in patients with gastric cancer.Methods:The medical records of gastric cancer patients undergoing surgery in Department of Gastrointestinal Surgery, Puji District, Dongguan People′s Hospital Affiliated to Southern Medical University from June 2018 to December 2019 were retrospectively analyzed, and 136 patients were selected.Collected patient data, including general information, the expression levels of tumor markers carcinoembryonic antigen(CEA), carbohydrate antigen 19-9(CA19-9), carbohydrate antigen 72-4(CA72-4), and the surgical method used.The relationship between the expression of tumor markers and radical resection was observed.Results:There was no statistically significant difference in the radical resection rate of gastric cancer in different genders, ages, and locations of gastric cancer (all P>0.05). TNM classification (tumor node metastasis classification) stage III patients had a lower radical resection rate (63.3%(38/60)) than stage I (100%(27/27)) and II (100%(49/49)) (χ 2=58.166 and 81.208, P<0.001). The radical resection rate of CEA(+ ) patients (47.8%(44/92)) was lower than that of CEA(-) patients (90.9%(40/44))(χ 2=23.394, P<0.001). The radical resection rate of CA19-9(+ ) patients (47.7%(42/88)) was lower than that of CA19-9(-) patients(87.5%(42/48))(χ 2=20.804, P<0.001). The radical resection rate of CA72-4(+ ) patients (54.3%(51/94)) was lower than that of CA72-4(-) patients (78.6%(33/42)) (χ 2=7.268, P=0.007). The variables with P<0.1 in univariate analysis were included in the logistic regression model, including 4 variables including TNM stage, CEA, CA19-9, and CA72-4.The results showed that TNM staging ( OR=1.169, 95% CI=0.925-1.634, P=0.001), CEA ( OR=1.067, 95% CI=1.364-4.338, P=0.024), CA19-9( OR=3.012, 95% CI=1.679-6.317, P=0.007), and CA72-4 were independent risk factors for radical resection( OR=5.364, 95% CI=0.675-3.224, P=0.004). The number of positive expressions of tumor markers was negatively correlated with the radical resection rate ( r=-0.621, P<0.05). The expression levels of CEA, CA19-9, and CA72-4 in patients with radical resection were 75.36(3.76, 198.20)μg/L, 152.76(34.81, 241.09)kU/L, 126.60(4.01, 218.07)kU/L, respectively.The expression levels of CEA, CA19-9, and CA72-4 in the radically resected patient group were 173.65(120.78, 254.87) μg/L, 255.88(102.45, 395.11) kU/L, 201.71(79.15, 325.92)kU/L.The expression level of CEA, CA19-9, CA72-4 in the group with radical resection were lower than that in the group with no radical resection, and the difference was statistically significant (the Z values were 10.672, 8.945, 9.862, all P<0.001). ROC curve showed that AUC of CEA, CA19-9 and CA72-4 were 0.627, 0.714 and 0.768, respectively.The best cut-off value of CA72-4 was 87.62 kU/L, the sensitivity was 88.1% (74/84) and the specificity was 90.4% (47/52). Conclusion:The expression levels and number of tumor markers CEA, CA19-9, CA72-4 in patients with gastric cancer were correlated with the risk of radical resection.
7.Effects of RNA interference targeting CDC25a gene on proliferation of human liver cancer HepG2 cells
Wei LI ; Ji CAO ; Lingli ZHOU ; Wang LUO ; Chun YANG ; Chengpiao LUO ; Yuan LI ; Jianjia SU
Chinese Journal of Pathophysiology 2014;(12):2142-2147
[ ABSTRACT] AIM:To investigate the effect of silencing cell division cycle 25a ( CDC25a) gene on the prolifera-tion of human hepatoma HepG2 cells.METHODS:CDC25a gene in human hepatoma HepG2 cells was silenced by RNA interference.Real-time PCR was applied to detect the expression of CDC25a, cyclin E and CDK2 at mRNA levels in the HepG2 cells.Western blotting was applied to detect the expression of CDC25a at protein level.In addition, MTT assay, Giemsa staining and flow cytometry were used to measure the proliferation of human hepatoma HepG2 cells.RESULTS:The expression of CDC25a at mRNA and protein levels in RNA silence group was lower than those in negative control group and normal control group (P<0.05).The mRNA expression of cyclin E and CDK2 in silence group was lower than that in negative control group and normal control group (P<0.05).The cell proliferation in silence group was lower than that in negative control group and normal control group ( P<0.05) .The results of flow cytometry revealed that the cells in silence group were blocked in G1 phase.CONCLUSION:Infection of LV-CDC25a-RNAi recombinant to the HepG2 cells effec-tively inhibits the CDC25a gene expression and the proliferation of human hepatoma cells, and arrests the cells in G1 phase, suggesting that CDC25a gene may be a key target for the treatment of liver cancer.
8.Effect of TanshinoneⅡA on Experimental Endometriosis in Rats
Lili ZHENG ; Wei LI ; Dou HUANG ; Lin LUO ; Xiaohong LUO ; Yuan LI
Herald of Medicine 2016;35(6):612-615
Objective To observe the effect of TanshinoneⅡA( TanⅡA) on endometriosis( EMS) rat model and study the possible mechanism. Methods EMS rat model was established by autologous endometrium transplantation. EMS rats were randomly divided into five groups: high-, medium-, low-dose TanⅡA group, mifepristone group and model control group( n=10), the other ten normal rats as normal control group.The rats in the model control group and normal control group were given 0.9% sodium chloride solution. The rats in the mifepristone group were given 25 mg . kg-1 . d-1 mifepristone. The rats in the high-, medium-, low-dose TanⅡA groups were given 30, 20 and 10 mg.kg-1.d-1 TanⅡA, respectively.Four weeks after the treatment, the volume of endometriotic implants in each rat was measured. Pathological changes of ectopic endometrium were observed under light microscope.Expression levels of Bcl-2, Bax and Caspase-9 in ectopic endomembrane of rats were determined by Western blotting. Results High and medium dose TanⅡA and mifepristone significantly reduced the volume of ectopic lesions(P<0.05).As compared with model control group, the inhibition rate of the ectopic lesions in low-, medium-, high-dose TanⅡA groups and mifepristone group was 17.81%, 57.78%, 58.88% and 64.12%, respectively.High and medium dose TanⅡA and mifepristone promoted atrophy of ectopic endometrium, inhibited the expression of Bcl-2 protein and promoted the expression of Bax and Caspase-9 in ectopic tissues( P<0.05) . Conclusion TanⅡA significantly decreases the size of ectopic lesions in rats, possibly through reducing the expression of Bcl-2 and increasing the expression of Bax and Caspase-9 to regulate cell apoptosis.
9.Detection of cell apoptosis induced by 125I brachytherapy in nude mice bearing lung adenocarcinoma using 99Tcm-Annexin V imaging combined with diffusion weighted MRI
Jinshan ZHANG ; Liangping LUO ; Changzheng SHI ; Yuan LI ; Wei AN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2014;34(5):385-389
Objective To detect apoptosis of transplanted lung adenocarcinoma cells in nude mice after 125I brachytherapy by 99Tcm-Annexin V combined with diffusion weighted MRI (MR-DWI).Methods Twenty-five BALB/c-nu nude mice models subcutaneously transplanted with A549 cells were divided into experimental group (EG,n=13) and control group (CG,n=12) by random number table method.One 125I seed with apparent activity of (24.8±6.3) MBq was implanted into each mouse in EG,while CG underwent cold seed implantation.Both of 99Tcm-Annexin V imaging and MR-DWI were performed within 7-10 d after brachytherapy,then all mice were sacrificed and tumor cell apoptosis was detected by terminal oxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) immunofluorescence,Survivin expression was assayed by SP.Two-sample t test,x2 test and Pearson correlation analysis were used for data analysis.Results Positive rate of cell apoptosis by 99Tcm-Annexin V imaging was 69.2%(9/13) and 8.3%(1/12) respectively in EG and CG (x2 =12.73,P<0.01).The uptake ratio of 99Tcm-Annexin V,apparent diffusion coefficient(ADC) value and apoptosis index(M) of the tumor in EG were 2.91±0.85,(2.03±0.44)×10-3 mm2/s and (49± 18) %,respectively,which were significantly higher than those of CG (1.26 ± 0.37,(1.29 ± 0.21) ×10-3 mm2/s and (11±4)% respectively,t=5.930,5.452,7.606,all P<0.05).Survivin expression in EG and CG was (46± 13) % and (15±7) % respectively (t =5.158,P<0.05).The value of ADC was correlated with AI and uptake ratio(r=0.756,0.788,both P<0.05).Uptake ratio was correlated with AI (r=0.754,P<0.05),while Survivin expression was negatively correlated with AI (r =-0.772,P<0.05).Conclusions Down-regulation of Survivin expression may play an important role in apoptosis induced by 125I brachytherapy.99Tcm-Annexin V combined with MR-DWI could effectively evaluate apoptosis of lung adenocarcinoma cells in a non-invasive way,thus it might be helpful in evaluation of early efficacy of 125I brachytherapy.
10.Effects of low-dose of TGF-β1 on maintaining bovine corneal stromal cell growth and retarding extra cellular matrix fibrosis in a three-dimensional culture model
Lu, ZHANG ; Shinan, LUO ; Jianbao, YUAN ; Weiwei, ZHOU ; Xia, LI
Chinese Journal of Experimental Ophthalmology 2017;35(5):396-403
Background Transforming growth factor-β1(TGF-β1) plays an important role in corneal woundhealing.The effects of TGF-β1 on the synthesis of extra cellular matrix (ECM) vary upon different concentrations.Previous studies focused on the effects of high concentration of TGF-β1 on keratocytes under the two-dimensional culture condition,and the effect of low concentration of TGF-β1 on the synthesis of ECM in keratocytes remains unclear.Objective This study was to investigate the growth of Pellet,a three-dimensional model of corneal stroma cells in vitro,and its ECM synthesis under a low concentration of TGF-β1.Methods Bovine corneal stromal cells were isolated from fresh bovine eyeballs by two-step digestion by collagenase and cultured using DMEM/F12 medium with 10% fetal bovine serum (FBS).Pellets derived fresh bovine keratocytes with culture medium containing 0.25 ng/ml TGF-β1 +5% FBS and 0.50 ng/ml TGF-β1 +5% FBS were established,respectively.The morphology of Pellets was observed under the natural light at 48 hours,1 week,2 weeks and 3 weeks after culture.In 3 weeks after culture,the cell structures was observed by hematoxylin-eosin staining,and Calcein-AM/propidium (Calcein-AM/PI) staining was used to assay the cell viability.Real-time flurorescence quantitative PCR and immunofluorescence technology were applied to analyze the expressions of α-smooth muscle actin (α-SMA),fibronectin (FN),type Ⅰ collagen (Col Ⅰ) and type Ⅲ collagen (Col Ⅲ) mRNA and proteins.RT-PCR was employed to detect the expressions of lumican (LUM) mRNA and keratocan (KERA) mRNA in the cells.Results Cells in Pellet clustered throughout the culture duration.Hematoxylin-eosin staining showed the mass red-dyed collagen fibers in both 0.25 ng/ml TGF-β1 +5% FBS group and 0.50 ng/ml TGF-β1 +5% FBS group,and most cells possessed complete structures.The death rate of the cells was (33.60±1.65)% in the 0.25 ng/ml TGF-β1 +5% FBS group and (30.90±0.78) % in the 0.50 ng/ml TGF-β1 +5% FBS group,showing an insignificant difference between them (t =0.144,P=0.887).The expressions of α-SMA,FN and Col Ⅲ proteins in 0.25 ng/ml TGF-β1 +5% FBS group were lower than those in the 0.50 ng/ml TGF-β1 + 5 % FBS group (tα-SMA =4.622,P =0.010;tFN =2.973,P =0.040;tCol Ⅲ =7.845,P<0.001),but the expression of Col Ⅰ in 0.25 ng/ml TGF-β1 +5% FBS group was higher than that in 0.50 ng/ml TGF-β1+5% FBS group (tColⅠ =4.022,P=0.016).The ratio of Col Ⅲ/Col Ⅰ in 0.25 ng/ml TGF-β1+5% FBS group was lower than that in the 0.50 ng/ml TGF-β1 +5% FBS group in both mRNA and protein level (tmRNA =-3.039,P =0.038;tprotein =3.215,P =0.032).The expression of LUM mRNA and KERA mRNA were detected in Pellet at different time points.The expression of LUM mRNA in 0.25 ng/ml TGF-β1 +5% FBS group increased over time.While in 0.50 ng/ml TGF-β1 +5% FBS group,the expression of LUM mRNA peaked at 1 week but declined at 2 weeks.The expression of KERA mRNA in two groups were all peaked at 1 week but declined at 2 weeks.Conclusions Low-dose TGF-β1 in Pellet can maintain the normal growth of keratocytes and synthesize ECM.The expression of ECM tends to the normal condition after reducing the concentration of TGF-β1,implying a scarless expression.