2.Preventive and therapeutical effects of N-acetylcysteine on selenite-induced cataract in vivo
Guang-ying, ZHENG ; Hua-jun, WANG ; Nan, TAN ; Gang, CHEN ; Yuan-yuan, LIANG
Chinese Journal of Experimental Ophthalmology 2012;30(6):515-519
Background The mechanism of both selenite-induced cataract and age-related cataract is oxidative damage.N-acetylcysteine (NAC) is one of the effective antioxidants,but the literature is little about the preventive and treating effects of NAC on cataract. Objective This study attempted to investigate the preventive and therapeutical effects of NAC on the selenite-induced cataract,and to discuss the possible mechanism. Methods Sixty 10-day-old clean SD rats were randomly divided into normal control group-1,normal control group-2,selenite-induced cataract group,NAC preventive group,NAC+normal saline group and NAC treatment group.Selenite cataract models were induced by subcutaneous injection of 3.46 mg/kg sodium selenite once daily for three days.The rats of NAC preventive group received the intraperitoneal injection of 2 mmol/L NAC 30 minutes before the injection of sodium selenite once daily for 6 days.In NAC treating group,2 mmol/L NAC was intraperitoneally injected 1 day after the injection of sodium selenite for 30 days,and the normal saline solution was injected at the same method in the NAC+normal saline group.Lens opacification was graded according to LOCS Ⅲ criteria.Histopathological change of lens epithelium was examined under a light microscope after hemotoxylin and eosin staining,and the ultrastructure was observed under the scanning electron microscope.The expression of caspase-3 in lens was assayed using immunochemistry.The levels of superoxide dismutase ( SOD ),malonaldehyde ( MDA ) in rat lens were detected respectively in corresponding time points.The use of the experimental animals complied with the Regulation for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Committee. Results In 7 days after experiment,lenses were completely clear in the normal control group.Lens opacification of Ⅴ grade was found in 11 eyes in selenite cataract model group,but no lens opacification of Ⅴ grade was seen in NAC preventive group,showing a significant difference(x2 =40.000,P<0.05 ).In 30 days after experiment,Ⅳ- Ⅴ grades of cataracts were found in 20 eyes both in NAC + normal saline group and NAC treating group (x2=0.153,P> 0.05 ).Histopathological examination showed that lens structure was normal,and the separation between LECs and anterior capsule,the rupture of cellular membrane,deformation of cellular nuclei and the feature of lens fiber were seen in selenite cataract group,but the damage of lens was mild in the NAC preventive group.Ultrastructure of lens was obviously abnormal in selenite cataract group,NAC+normal saline group and NAC treating group.Expressions of caspase-3 and SOD in lens were significantly lower,but that of MDA was significantly higher in the selenite cataract group than the normal control group (P<0.05) ;while those of the NAC preventive group were significantly different from selenite cataract group(P<0.05).No significant difference was found in the expressions of caspase-3 and the levels of SOD and MDA between NAC+normal saline group and NAC treating group (P>0.05 ). Conclusions Selenite can induce the apoptosis of LECs.NAC can evidently postpone formation of selenite cataract by increasing the activity of SOD,decreasing the level of MDA and the expression of caspase-3.However,NAC could not reverse selenite-induced lens damage.
3.Recent advances of MDM2-p53 inhibitors
Ji-chao WANG ; Cai-bin YANG ; Yi-lin ZHUO ; Chong LIANG ; Jun-gang WANG ; Sheng DONG ; Bo-teng LI ; Shu-hua ZHANG ; Guo-gang ZHANG
Acta Pharmaceutica Sinica 2021;56(4):983-995
Protein-protein interactions (PPI) are involved in a variety of biological processes, including cell-to-cell interactions, metabolism and development control. The misregulation, post-translational modification and interference of PPI are related to a variety of human diseases, making the regulation of these interactions a very attractive field of drug discovery. In recent years, the interaction between MDM2 and p53 has become a research hotspot, which plays an important role in the treatment of tumors. But unfortunately there are no such inhibitors approved all over the world. In this view, recent advances of MDM2-p53 inhibitors were briefly described and its inhibitors with potential therapeutic activities in clinical studies were introduced.
4.Feasibility of cartilage link protein of hyaluronic acid for defining radiotherapeutic target volume in a mouse model of lung tumor.
Zhi-Xin LIANG ; Yong-Gang QIANG ; Yong-Hua LIAO
Journal of Southern Medical University 2012;32(3):301-305
OBJECTIVETo investigate the feasibility of using cartilage link protein of hyaluronic acid (HA-CLP) for defining the tumor boundary in a mouse model of lung carcinoma.
METHODSLung carcinoma was induced in KM mice by chemical carcinogenesis. HA-CLP separated from bovine cartilage and purified by affinity chromatography was labeled with (125)I for autoradiography. Immunohistochemical analysis and Western blotting were used to examine the efficiency of HA-CLP in defining the boundaries of the lung tumors.
RESULTSWith autoradiography, the clearest image of lung cancer was obtained at 2 h. With immunohistochemical method, the tumor boundary was the most clearly displayed at 2 h when the strongest signals of HA-CLP was detected; Western blotting also showed the clearest bands of HA-CLP at 2 h.
CONCLUSIONHA-CLP has the immunogenicity of HABP, and can efficiently indicate lung tumor boundary in autoradiography and immunohistochemistry.
Animals ; Autoradiography ; methods ; Extracellular Matrix Proteins ; metabolism ; pharmacology ; Female ; Hyaluronic Acid ; metabolism ; Immunohistochemistry ; Iodine Radioisotopes ; Lung Neoplasms ; radiotherapy ; Male ; Mice ; Proteoglycans ; metabolism ; pharmacology ; Radiotherapy, Image-Guided ; methods
5.Protective effect of dexamethasone on stress response induced by intrauterine balloon aortic valvuloplasty in a fetal lamb model
Kaiyu ZHOU ; Yimin HUA ; Liang ZHAO ; Yifei LI ; Chuan WANG ; Ming YANG ; Hongyu DUAN ; Gang WU ; Nan GUO ; Qi ZHU
Journal of Clinical Pediatrics 2013;(7):660-665
Objective To investigate the protective effect of dexamethasone on stress response induced by intraute-rine balloon aortic valvuloplasty (IUBAV) in a fetal lamb model. Methods Twenty-four near term twin pregnant goats were randomly assigned to control group (n=12) and dexamethasone group (intracardiac injection of dexamethasone) (n=12). Ultrasound guided IUBAV model was established. According to the combination of treatment and operation, 48 fetal lambs were divided into four groups. The following parameters were dynamically monitored by ultrasound, which were fetal and neonatal heart rate (HR), cardiac rhythm, aortic resistance index (RI) and pulsatility index (PI). On established IUBAV model, blood samples and liver tissue specimens were taken from fetal/neonatal goats for detection of blood glucose (Glu), lactate acid (LA), plasma epinephrine (E), norepinephrine (NE), cortisol (Cort), hepatic glycogen staining. Results IUBAV resulted in increased values of Glu, LA, E, NE and Cort levels, and decreased pH value, there were significant differences between pre-and 3h-post procedure (P<0.05), and significant differences were also existed in these values between pre-and 3d-post procedure (P<0.05). After administrating dexamethasone, the changes of Glu, LA, E, NE, Cort and pH levels were suppressed effectively. Fetal hepatic glycogen was consumed in large amounts due to IUBAV while recovered 3 days after IUBAV by glycogen staining. After administrating dexamethasone, hepatic glycogen consumption related to IUBAV was obviously inhibited. After IUBAV, fetal aortic RI was increased, and there was significant differences compared with pre-procedure (P<0.05). Up to 3d-post procedure, the values of RI recovered to some extent, but statistical difference was exist-ed compared with pre-procedure (P<0.05). After administrating of dexamethasone, increased aortic RI was effectively sup-pressed. Conclusions IUBAV could lead to reversible stress response and increased aortic RI in a fetal lamb model which could be alleviated by dexamethasone.
6.Effects of thyroid hormone on the expression of homeobox gene Nkx6.1 in the cerebrum tissue of offspring of hypothyroidism rat
Ren, NA ; Rui, ZHANG ; Bei-lei, WANG ; Jing-hua, LI ; Yuan, LI ; Dong-chun, LIANG ; Gang, GUO
Chinese Journal of Endemiology 2010;29(2):150-154
Objective To explore the effects of thyroid hormone on the expression of homeobox gene Nkx6.1 in offspring of hypothyroidism rats and the relationship between gene expression and hormone level by supplying their hypothyroidism pregnant mother with thyroid hormone. Method A total of 240 Wistar rats were half nude and half female. Female rats were randomly divided into eight groups: control, hypothyroidism group, hypothyroidism groups which were supplied with thyroid hormone in high, medium and low dosage in early stage(1- 17 d) and in late stage( 18 - 20 d). According to 100 grams of body weight, the concentration of thyroid hormone were 3.5,2.0,0.5 μg/d in high, medium and low dosage group. All the rats were fed with low-iodine food. The normal control group was given KIO_3 solution and the other groups were given deionized water. After three months female rats were mated with male rats. The content of Nkx6.1 mRNA in brain tissue of 17-day fetal rats, new-born and 20- day old offspring by real-time fluorescence quantitative PCR techniques. Results①A rat model of hypothyroidism was successfully established, there were statistical significance between 8 groups in TT_3,TT_4,FT_3,FT_4(F=4.08,31.99,5.79,26.34, all P < 0.01 ). ② The expression of Nkx6.1 mRNA had significant difference(F = 758.720, 1121.589,144.716, all P < 0.01 ) between groups in 17-day fetal rats, new-bern and 20-day old offsprings and intra- groups in different time (F=2898.863,325.605,716.285,56.329,236.727,196.678,7115.752,9152.306, all P < 0.01 ). ③The time factor and dosage factor had influence on Nkx6.1 mRNA expression(F = 1176.655,246.530, all P < 0.01 ). There were interaction between time and dosage factor(F = 1249.934, P < 0.01 ). ④Comparison of Nkx6.1 mRNA expression between hypothyroidism group and normal control group had significant difference in the above three time points(all P < 0.01 ). ⑤Comparisons of Nkx6.1 mRNA expression between 6 hypothyroidism groups which were supplied with thyroid hormone and hypothyroidism group had significant difference(all P < 0.01 ) in new-bern and 20-day old offspring; comparisons of Nkx6.1 mRNA expression between hypothyroidism groups which were supplied with high and medium thyroid hormone and hypothyroidism group had significant difference in 17-day fetal rats(all P < 0.01 ). ⑥Comparison of Nkx6.1 mRNA expression between hypothyroidism groups which were supplied with medium thyroid hormone in early stage and normal control group had no statistical significance (all P > 0.05), while between the other 5 groups which were supplied with thyroid hormone and normal control group had significant difference(all P < 0.01 ) in the above three time points.⑦Multiple comparison of early stage groups which were supplied with thyroid hormone showed that the expression of Nkx6.1 mRNA had significant difference(all P < 0.01) between high, low dosage groups and medium group in 17-day fetal rats, new-bern and 20-day offspring(all P< 0.01). ⑧Multiple comparison of late stage groups supplied with thyroid hormone showed that old offspring and between high dosage groups and low dosage groups in 17-day fetal rats and 20-day the expression of Nkx6.1 mRNA had significant difference(all P < 0.01 ) between three groups in new-bern and 20- day old offspring. Conclusion The expression of Nkx6.1 in rats offspring is highly related to the supply dosage and supply time of thyroid hormone in hypothyroidism pregnant rats.
7.The effects of intergrin-linked kinase on angiogenesis in hypertrophic scar.
Ren-Kun WANG ; Ye-Yang LI ; Gang LI ; Wei-Hua LIN ; Jing-En SUN ; Zhen-Wen LIANG ; Xiao-Hong WANG
Chinese Journal of Plastic Surgery 2013;29(6):413-412
OBJECTIVETo investigate the effects and regulatory mechanism of ILK on angiogenesis in hypertrophic scar.
METHODSThe human scar microvascular endothelial cells (HSMECs) were isolated from 6 patients' hypertrophic scar in vitro. The HSMECs with good condition in 2nd to 4th generation were selected as experimental objectives. (1) HSMECs were divided into the blank control group (treated with routine culture), negative control group (treated with only Lipofectamine 2000), LY294002 group (incubated with 50 nmol/L LY294002), ILK siRNA group (incubated with 20 nmol/L ILK siRNA). RT-PCR and Western Blot were used to detect the expression of ILK mRNA and its protein after transfecion for 48 h. (2) The digested HSMECs of four groups were resuspended with DMEM without serum and then seeded onto the upper compartment of transwell insert which contained complete medium in its lower compartment. The cell migration experiment was stopped in 10 h and then the migrated cells were counted to analyze the effects of different interventions on the migration ability of HSMECs. (3) The thawed ECMatrix was put into each well of pre-colled 48-well tissue culture plate, and then the plate was put into the incubator at 37 degrees C to make it to become gel. The HSMECs of four groups were seeded onto the surface of the ECMatrix gel and were put into incubator. Eight random view-fields per well should be valued by the sheet of pattern recognition about angiogenesis after 8 hours to evaluate the ability of angiogenesis in vitro between four groups.
RESULTS(1) The expression of ILK mRNA (ILK mRNA = 0.829 +/- 0.109, t = 13.151, P = 0.006) and protein (ILK protein = 0.096 +/- 0.049, t = 36.656, P = 0.000) were both inhibited obviously in ILK siRNA group compared with the blank control group (ILK mRNA = 0.829 +/- 0.109, ILK protein = 1). And, the expression of ILK in LY294002 group was slightly lower than that of black control group, but there was no statistical difference. (2) The number of migrated cells in ILK siRNA group (88.111 +/- 3.079) and LY294002 group (138. 667 +/- 2.404) were respectively lower than that in blank control group (322.333 +/- 3.712, P < 0. 05) in 10th hour. (3) Compared to blank control group (4.333 +/- 0.191), the ability of angiogenesis in vitro decreased significantly ILK siRNA group (2.625 +/- 0.125) and LY294002 group (3.125 +/- 0.250), in which, the vascular network structures were not formed perfectly in 8th hour (P < 0.05).
CONCLUSIONSThe ability of HSMECs' migration and angiogenesis in vitro are inhibited significantly when the expression of ILK is down-regulated. It reveals that ILK may play an role in the regulation of scar angiogenesis.
Cell Movement ; Cell Proliferation ; Chromones ; pharmacology ; Cicatrix, Hypertrophic ; enzymology ; pathology ; Endothelial Cells ; cytology ; drug effects ; Humans ; Lipids ; pharmacology ; Morpholines ; pharmacology ; Neovascularization, Pathologic ; etiology ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; physiology ; RNA, Messenger ; analysis ; RNA, Small Interfering ; metabolism
8.Expression of TGF-β1 protein in orbital bone after enucleation with placement of an orbital implant
Jian-min, MA ; Jia-liang, ZHAO ; Zhi-hui, LI ; Feng-hua, CHEN ; Ji-tong, SHI ; Gang-wei, CHENG ; Xin, GE
Chinese Journal of Experimental Ophthalmology 2011;29(6):517-520
Background Placement of an orbital implant is a main way to prevent orbital atrophy with aging.But its mechanism is under clear.Researchs showed that bone growth factors play important role during the development and repair of bone,especially transforming growth factor-β1(TGF-β1).Objective Present study was to investigate the expression of TGF-β1 protein in orbital bone after enucleation or enucleation with placement of an orbital implant and its function in the mechanisms of preventing and treating the orbital malformed development after enucleation with placement of an orbital implant.Methods Twenty-one age- and weight-matched New Zealand white young rabbits were randomly divided into the enucleation,implant and control groups,and each group including seven rabbits.Eyeball nucleation surgery was performed in the left eyes of 7 1-month-old rabbits,and a spherical orbital implant was inserted after enucleation of the left in matched rabbits in implant group.The left eye of normal rabbits served as controls.The rabbits were sacrificed in 1 month after surgery.The expression of TGF-β1 protein in the left orbital bone was detected using enzyme immunoassay and FITC labelling immunoassay technique in the sections of zygomatic bones.The content of TGF-β1 protein in the left orbital bone tissue was measured by ELISA method.This use of animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The height and width of orbital in enucleation group were significantly lower than those of implant and normal control groups(height:P=0.00,P=0.00;width:P =0.00,P=0.00).The positive bone cells of both enzyme immunoassay and FITC staining were increased in the implant and control groups in comparison with enucleation group,but the positive response intensity for TGF-β1 was resembled between implant group and control group.ELISA result revealed that the content of TGF-β1 protein in bone tissue was significantly lower in the enucleation group than in implant and control groups(P=0.00,P=0.00).The expression and content of TGF-β1 protein in bone tissue is similar between the implant group and the control group(P=0.41). Conclusion The experiment results indicate that TGF-β1 protein participate in the orbital development.TGF-β1 played important role in the prevention and treatment of enucleation-induced orbital malformation in the eye with placement of an orbital implant.
9.Effects of hyperthyroidism on the electrophysiological characteristics between atrium and gap junction between atrium and pulmonary vein
Liang-Rong ZHENG ; Qing-Gang ZHANG ; Jun-Zhu CHEN ; Jun-Xian SONG ; Jian-Hua ZHU ; Jian SUN ;
Chinese Journal of Emergency Medicine 2006;0(12):-
Objective To investigate the effects of hyperthyroidism on the electrophysiological characteristics of atrium and gap junction between atrium and pulmonary vein.Methods Twenty-four rabbits were randomly divided into control group and hyperthyroid group.Atrium and pulmonary vein were dissected after the atrial effective refractory period (AERP)was measured.Connexin 43(Cx43)and Counexin 40(Cx40)protein and mRNA were determined by Western blot and semi-quantitative reverse transcription-polymerase chain reaction,respectively.Results In comparison with control group,AERP and rate adaption of AERP in hyperthyroid group were significantly shorter than that of control group( P<0.01).The concentration of Cx43 protein in hyperthyroid group was significantly higher than that of control group,but Cx40 protein was significantly lower than that of control group(P<0.05).The expression of Cx43 mRNA in atrium and pulmonary vein was found to be up-regulated in hyperthyroid group as compared with that of control group(P<0.01). With the level of Cx40 mRNA,there was no significant difference between two groups.Conclusion Thyroid hormone could lead to remodeling of both atrial electrophysiology and gap junction between atrium and pulmonary vein.
10.Short and long-term efficacy of focused ultrasound therapy for vulva dystrophy
Cheng-Zhi LI ; Du-Hong BIAN ; Lian WANG ; Hua-Qin WANG ; You-De CAO ; Nian-Gang YIN ; Chun-Liang ZHAO ;
Chinese Journal of Obstetrics and Gynecology 2001;0(01):-
0.05).However,the shorter the history of the disease,the better the efficacy of the treatment.The younger the patient was,the better the efficacy of the treatment.Conclusions Vulva dystrophy can be treated with focused ultrasound effectively and safely.This approach appears to be a new promising treatment method.