1.Effects of glycyrrhizin-18? on expressions of CTGF and TGF-?1 in renal interstitium in rats with obstructive nephropathy
Kailong LI ; Jianguo ZHANG ; Huimin WANG ; Jin CHEN
Journal of Third Military Medical University 1983;0(04):-
Objective To investigate the effects of glycyrrhizin-18? on the expressions of connective tissue growth factor (CTGF) and transforming growth factor-?1 (TGF-?1) in renal interstitium in rats with obstructive nephropathy. Methods A total of 100 Wistar rats were divided into experiment group (include prophylaxis group with glycyrrhizin-18?, glycyrrhizin-18? treatment group, and group with normal saline) and control group (sham operation group). Models of rats with obstructive nephropathy were established by unilateral ureteral ligation. The rats were sacrificed at 7, 14, 28, and 56 d after operation and specimens were taken from the renal cortex. The morphopathological changes in the renal interstitium were observed byr light and electron microscopy. CTGF, TGF-?1, and collagen type Ⅲ were detected by immunohistochemical staining. The measured data were evaluated by EIG image analysis system. Results In the renal interstitium, there was progressive fibrosis in the model. Semi-quantitative analysis indicated that the expression levels of CTGF, TGF-?1, and collagen type Ⅲ in the renal interstitium in glycyrrhizin-18? treatment and prophylaxis groups were markedly higher than those in the sham operation group (P0.05). The expression of CTGF in the renal interstitium was closely correlated with those of TGF-?1 and collagen type Ⅲ (P
2.Effect of castration on miR-301a expression in a mouse xenograft model of human prostate cancer
Kailong LIU ; Li WANG ; Yong ZHANG ; Haitao GAO ; Xiaolu WANG ; Jinchun QI ; Wei LI
Chinese Journal of Urology 2013;(2):135-139
Objective To investigate the changes of miR-301a and its host gene expression SKA2 in LNCaP prostate cancer xenografts in the castrated nude mice.Methods LNCaP cells were inoculated subcutaneously in nude mice to establish xenograft models of human prostate cancer.When the tumor volume grew to 200 mm3,the nude mice were randomly divided into the following 4 groups(n =6): 2 groups of nude mice to surgical castration,the other 2 groups of mice as control groups.The growth of those xnografts in nude mice was observed weekly and a growth curve of the xenografts was drawn.A point time during the process of tumor-regressing was selected when a group of castrated nude mice and a group of control mice were killed(the first time).The other 2 groups nude mice were continued to be observed.Another point time in the process of tumor re-growth,the rest castrated nude mice and control mice were killed(the second time).The tumors were weighted and the inhibitory rate was calculated.MiR-301a and SKA2 expression were detected by real-time PCR.Results The growth of the xenografts gradually decreased in LNCaP xenografts in nude mice after castration.After 13 days,the xenografls sizes decreased to(62.5 ±21.5)mm3 and tumor inhibitory rate was 59.8%(t =-3.895,P =0.018)in castration group of nude mice.At the 17th day after castration,tumor volume reached the minimum,and then gradually increased.At the 41st day after castration,tumor volumes in castration group increased to(364.5 ±97.3)mm3 and the tumor inhibitory rate was 62.2%(t =-7.017,P =0.002).MiR-301a and SKA2 in the first time of xenografts from the castrated group were both significantly lower than those of the control group(0.65-fold and 0.50-fold,respectively).However,their expressions in the second time of xenografts from the castrated group increased and were consistent with the control group(P > 0.05).Conclusions Castration could turn prostate cancer xenografts from androgen-dependent into androgen-independent.There could be a close correlation between the characteristic of prostate cancer androgen-dependent and the expressions of miR-301 a and host gene SKA2.
3.Effects of endoplasmic reticulum stress on albumin-induced apoptosis in renal proximal tubular cells
Xiaowei WU ; Yani HE ; Huiming WANG ; Kailong LI ; Hanlu DING ; Shunqiao LV
Chinese Journal of Nephrology 2009;25(3):198-203
ObjectiveTo investigate the effects and molecular mechanism of endoplasmic reticalam stress (ERS) on albumin-induced apoptosis in renal proximal tubular cells (HKCs). MethodsWestern blot was performed to detect the relationship of the expression of glucose-regulated protein 78 (GRF78) and CCAAT/enhancer-binding protein-homologous protein (CHOP) with the action time and concentration of haman serum albumin (HSA). Expression levels of CHPO mRNA and protein in HKCs after CHOP siRNA transfection were examined by real-time fluorescence quantitative PCR and Western blot respectively. Annexin-V-FITC and PI doable staining cytometry was used to detect the apoptosis of HKCs induced by HSA and influenced by CHOP siRNA. Results(1)After HKCs were stimulatde by 0, 5, 10, 20 g/L albumin for 24 hours respectively, the expression of GRP78, CHOP and HKCs apoptosis were increased with the albumin concentration (P<0.01). After HKCs were stimulated by 20 g/L albumin for 0, 6, 12, 24, 36 hours respectively, the expression of GRP78 was up-regulated at 6-hour, while CHOP and HKCs apoptosis were increased at 12-hour, and significant differences were found among groups (P<0.01). (2) CHOP siRNA significantly inhibited albumin-induced HKC CHOP mRNA and protein expression, as well as HKC apoptosis (P<0.01). ConclusionsRenal tubular cells exposed to high protein load result in EBS. ERS may subsequently lead to tubular damage by activation of pro-apoptosis factor CHOP.
4.Nuclear factor- κB activation is involved in MCP- 1 expression in human mesangial cells induced by Ox- LDL
Huiming WANG ; Miaozhen ZHU ; Xiang XU ; Jianguo ZHANG ; Yani HE ; Kailong LI
Chinese Journal of Pathophysiology 2005;21(11):2214-2220
AIM: To investigate the role of nuclear factor- κB (NF- κB) in the expression of monocyte chemoatractant protein- 1 (MCP- 1) in human mesangial cells (HMCs) induced by oxidized low- density lipoprotein (Ox- LDL).METHODS: HMCs were used as target cells. Inhibitory κBα (IκBα) and MCP- 1 protein level was measured by cell ELISA.Activities of transcriptional factors NF- κB were determined by electrophoresis mobility shift assay (EMSA). Immunohistochemistry was used to detect the translocation of Rel p65. RESULTS: NF - κB DNA - binding activation in MCs was observed when 10-100 mg/L Ox - LDL was added to the medium, and 50 mg/L Ox - LDL caused the strongest effect (8.50 ± 1.14, P < 0.01vs control; P < 0.05 vs 10, 25 and 100 mg/L Ox - LDL). The most optimal stimulation time was 60 min ( 11.0 ± 2.11, P <0.01 vs control; P < 0.05 vs 30 min or 240 min). IκBα protein level in MC dropped down most obviously after 60 min incubation with 50 mg/L Ox - LDL (0.050 ± 0.006, n = 5, P < 0.01 vs control), while MCP- 1 expression level was the highest (0.331± 0.016, n = 5, P < 0.01 vs control). The translocation of Rel p65 from cytoplasm to nucleus was detected too. NF - κB inhibitor pyrroledithiocarbomate (PDTC) could inhibit these effects induced by Ox- DL. CONCLUSION: Activation of NF- κB regulate the expression of MCP- 1 in HMCs induced by Ox - LDL.
5.Protective role of glycyrrhizin on experimental obstructive nephropat hy in rats
Kailong LI ; Kunxia JIA ; Jianguo ZHANG ; Huimin WANG ; Jing CHEN ; Guoping WU
Journal of Third Military Medical University 2001;23(5):573-575
Objective To investigate the protective role of glycyrrhizin on experimental obstructive nephropathy in rats. Methods The model of obstructive nephropathy was induced by unilateral uretera l ligation in rats and the animals were sacrificed at 12 h and 3,14,28,56 d resp ectively after operation. Specimens were taken from the cortex of kidney. In re nal stroma, routine morphological examinations and counts of karyocyte were made , the expression of collagen type Ⅰand type Ⅲ were also detected by immunohist ochemical staining,then, semi-quantified by EIG image analysis system. Results A progressive fibrosis was observed in renal stroma of the mod el. 12 h after operation, counts of karyocyte increased markedly i n renal stroma in groups of treatment and pretreatment with glycyrrhizin compar ed with that in sham-operation group(P<0.05),and decreased distinctly compared with that in saline treated group(P<0.01), but no significant di fference was found between treatment group and pretreatment group(P>0.05) ;3 d after operation, in group treated or pretreated with glycyrrhizin. The e xpression of collagen type Ⅰ increased markedly compared with that in sham-o peration group(P<0.01) in renal stroma, but decreased notably compare d with that in saline treated group(P<0.01),while, there was not any diffe rence between treatment group and pretreatment group(P>0.05). The expressi on of collagen type Ⅲ in renal stroma was almost the same as that of coll agen type Ⅰ. Conclusion Glycyrrhizin has some therapeutical bu t no preventive effect to experimental obstructive nephropathy in rats.
6.Nuclear factor-?B activation is involved in MCP-1 expression in human mesangial cells induced by Ox-LDL
Huiming WANG ; Miaozhen ZHU ; Xiang XU ; Jianguo ZHANG ; Yani HE ; Kailong LI
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: To investigate the role of nuclear factor-?B (NF-?B) in the expression of monocyte chemoatractant protein-1 (MCP-1) in human mesangial cells (HMCs) induced by oxidized low-density lipoprotein (Ox-LDL). METHODS: HMCs were used as target cells. Inhibitory ?B? (I?B?) and MCP-1 protein level was measured by cell ELISA. Activities of transcriptional factors NF-?B were determined by electrophoresis mobility shift assay (EMSA). Immunohistochemistry was used to detect the translocation of Rel p65. RESULTS: NF-?B DNA-binding activation in MCs was observed when 10-100 mg/L Ox-LDL was added to the medium, and 50 mg/L Ox-LDL caused the strongest effect (8.50?1.14, P
7.Influence of albumin-activated renal proximal tubular epithelial cells on peritubular capillaries and its potential mechanism
Jing LIN ; Yani HE ; Huiming WANG ; Kailong LI ; Hanlu DING ; Haiying SHEN ; Jvrong YANG
Chinese Journal of Nephrology 2008;24(9):657-662
Objective To explore the influence of albumin-activated renal tubular epithelial cells (RTECs)on peritubular capillaries in co-culture system and its potential mechanism. Methods Endocytosis of TRITC labeled bovine scrum albumin (TRITC-BSA) by HKC was detected by laser scanning confocal fluorescence microscope. HKC or HKC transfected with cubilin (endocytic receptor of albumin) siRNA or pre-treated with rotenone was incubated with albumin(20 g/L) for 24 h respectively. Fluorescence probe technique and spectrometry were applied for determination of intracellular superoxide anion O2-and H2O2 in supematant. Then, the albumin-aetivated-HKC, pretreated-HKC with cubilin siRNA or rotenone, was cultured with HUVEC for 24 h in co-culture system respectively. HUVEC proliferation was determined by MTT and cellular apoptosis was analyzed by flow cytometry. Tabular morphogenesis of endothelial cells was examinedby microscopy. Results TRITC-BSA uptake was obviously lower in HKC transfected with cubilin siRNA. Intracellular generation of O2-and H2O2 in culture supernatant was increased in dose-and time-dependent manner after stimulating with albumin. The levels of O2-and H2O2 were suppressed by cubilin siRNA and rotenone. In co-culture system, albumin-activated-HKC induced endothelial cells apoptosis and inhibited their capillary tubular morphogenesis. Pretreatment of HKC with cubilin siRNA or rotenone could suppress endothelial cells apoptosis and promote capillary tubular morphogenesis. Conclusions There may be a crosstalk between RTECs and peritubular microvascular endothelial cells in renal proteinurie diseases. The generation of ROS by albumin-activated RTECs may play an important role in this process.
8.Efficacy of hirudin in treating immunoglobulin A nephropathy with hematuria: a randomized controlled trial
Kailong LI ; Yani HE ; Hongwei ZUO ; Huimin WANG ; Hanlu DING ; Jurong YANG
Journal of Integrative Medicine 2008;6(3):253-7
OBJECTIVE: To investigate the curative effects and side effects of hirudin in treating immunoglobulin A nephropathy (IgAN) with hematuria and minimal proteinuria in a short-term. METHODS: Two hundred and sixty-two histologically confirmed cases of IgAN with hematuria and minimal proteinuria from 1998 to 2007 were randomly divided into hirudin-treated group (peroral administration of Maixuekang capsules) and dipyridamole-treated group (peroral administration of dipyridamole). In the two groups, contrast analysis of conformation and counts of erythrocytes in urine, urine protein quantitation in 24 hours, levels of serum creatinine (Scr) and creatinine clearance rate (Ccr), blood lipid, five items of blood clotting and side effects was performed. RESULTS: After six-month treatment, the anisotrophy rate and the counts of erythrocytes in urine, and the urine protein quantitation in 24 hours in hirudin-treated group were decreased distinctly as compared with pre-treatment (P<0.01) and dipyridamole-treated group (P<0.05). On the other hand, Ccr was increased obviously in hirudin-treated group as compared with pre-treatment and dipyridamole-treated group (P<0.01). The blood lipid was also ameliorated in hirudin-treated group, but there was no significant difference. The anticoagulation effect of hirudin was better than dipyridamole (P<0.01). Efficacy assessment showed that the total response rate, complete remission rate and predominance remission rate in hirudin-treated group were higher than those in dipyridamole-treated group. Few side effects were found in both groups, and the rate of adverse reaction in gastrointestinal tract was lower in hirudin-treated group as compared with that in dipyridamole-treated group (P<0.05). CONCLUSION: Compared with dipyridamole, hirudin has superiority in kidney protection and decreasing the anisotrophy rate, counts of erythrocytes in urine and the urine protein.
9.Clinical study of venous phase CT value in predicting central group lymph node metastasis of papillary thyroid carcinoma
Jinhao LI ; Guidong WANG ; Xuefei LI ; Zilin LIU ; Kailong MENG
Journal of International Oncology 2022;49(10):581-585
Objective:To investigate the clinical value of imaging features of primary lesions combined with venous phase CT value in predicting central group lymph node (LN) metastasis in patients with papillary thyroid carcinoma (PTC) .Methods:Clinical data of 170 PTC patients who underwent central group LN dissection in the First People's Hospital of Handan City of Hebei Province from January 2017 to June 2020 were retrospectively analyzed. All patients were divided into different groups according to whether central group LN metastasis or not, and there were 89 patients with central group LN metastasis and 81 patients without central group LN metastasis. The CT value and imaging features of primary lesions in different periods were analyzed, and the imaging features of primary lesions combined with venous phase CT values to predict the central group LN metastasis were evaluated by the receiver operating characteristic (ROC) curve.Results:There were no statistically significant differences in CT value in plain scan phase and CT value, net increased CT value, standardized CT value in arterial phase between patients with and without central group LN metastasis (all P>0.05) . The CT value, net increased CT value and standardized CT value in venous phase of patients with central group LN metastasis were (113.84±22.95) HU, (59.05±12.10) HU and 0.72±0.14 respectively, which were significantly higher than those of patients without central group LN metastasis [ (103.99±17.67) HU, (51.29±14.45) HU and 0.59±0.10] ( t=3.26, P<0.001; t=3.81, P<0.001; t=3.67, P<0.001) . ROC curve analysis showed that the area under the curve for diagnosing central group LN metastasis of PTC patients was 0.75, 0.70 and 0.76 when the cut-off values of CT value, net increased CT value and standardized CT value in venous phase were 115.78 HU, 62.37 HU and 0.75 respectively. There were statistically significant differences in the diameter of primary focus and the contact area of thyroid capsule between patients with and without central group LN metastasis ( Z=-2.34, P=0.019; Z=-2.08, P=0.037) . There were no statistically significant differences between calcification and primary lesion location (both P>0.05) . Lesion diameter >2 cm (87.73%) and capsule contact range ≥1/2 (92.17%) had the highest specificity in predicting central group LN metastasis. The imaging features of primary lesion combined with standardized CT value in venous phase was in good agreement with histopathological diagnosis results in predicting central group LN metastasis (Kappa=0.475) , and the sensitivity and specificity were 73.12% and 82.75% respectively. Conclusion:The imaging features of the primary lesion combined with CT value in venous phase have a good clinical value in predicting central group LN metastasis in PTC patients. Patients with primary lesion diameter >2 cm, capsule contact range ≥1/2 and the standardized CT value in venous phase >0.75 are more likely to have central group LN metastasis.
10.The design and application of a moxibustion instrument with less harmful smoke and direction adjustment.
Hongliang TANG ; Xiongjiang WANG ; Zhangsong NONG ; Kailong WANG ; Jun PANG
Chinese Acupuncture & Moxibustion 2018;38(2):217-221
The design of a clip-on moxibustion instrument which could adjust the directions and absorb harmful granules of moxa smoke was introduced in this paper. It was designed to solve the problems in home health care and clinical treatment, such as the inconvenience of adjustment of moxibustion directions and temperature, more dust and granules of moxa smoke and inconvenience of moxibustion placement. The main part of moxibustion instrument was made up of moxibustion box, bracket and base clamp. The moxibustion box could fix moxa stick and absorb smoke granules; the bracket could be twisted to adjust the direction of moxibustion box; the base clamp was aimed to fix moxibustion box on the edge of the object to save space. This moxibustion instrument could be used for various indications of moxibustion, especially in the joints or body parts with less muscles; and it can significantly enhance the pertinence and safety of moxibustion, and reduce the labor intensity. This moxibustion instrument is original and unique, simple and reasonable, easy to operate, with low cost and good effect, which will increase new vitality for the popularization and development of moxibustion.