Objective:This study aimed to investigate the effects and the molecular mechanism of JNJ-7706621 on cell cycle and apoptosis of breast cancer cell lines. Methods:Breast cancer cell lines T47D and MDA-MB-231 were cultured in vitro and treated with JNJ-7706621 at varying concentrations. MTT assay was used to measure cell proliferation. Flow cytometry was applied to analyze the distribution of cell cycle and apoptotic rates. Western blot was performed to analyze the expression of cyclin B1-CDK, the phosphoryla-tion levels of CDK1Thr161 and CDK1Tyr15, as well as the expression of p53, Bcl-2, caspase3 and poly(ADP-ribose) polymerase (PARP). Results:JNJ-7706621 inhibited the proliferation of the T47D and MDA-MB-231 cells in a dose-and time-dependent manner. Flow cytometry showed that the T47D and the MDA-MB-231 cells in the G2/M-phase significantly increased after treatment at varying concentrations (0, 1, 2, 4μM) of JNJ-7706621:the G2/M-phase rates at the corresponding concentrations were (12.66±1.55)%, (20.63± 1.32)%, (23.20±1.82)%, and (32.19±2.37)%, respectively, for the T47D cells (P<0.05), and the G2/M-phase rates were (16.22±1.48)%, (21.45±0.85)%, (25.25±1.26)%, and (31.08±1.16)%, respectively, for the MDA-MB-231 cells (P<0.05). The rates of apoptotic cells were also significantly increased (P<0.05). Western blot results indicated that JNJ-7706621 exerted a slight effect on the expression of CDK1 and p53 and that the phosphorylation level of CDK1 decreased at the Thr161 site but increased at the Tyr15 site. In addition, cleaved caspase3 and PARP increased, whereas Bcl-2 and cyclinB1 decreased significantly. Conclusion:JNJ-7706621 can significantly inhibit the proliferation of breast cancer cell lines T47D and MDA-MB-231 by inducing cell cycle arrest and apoptosis, which may be associated with the downregulation of the CDK1 phosphorylation at the Thr161 site.

Objective To assess the effects of medicated permeation with traditional Chinese medicine Shujin-Zhitong liquid combined with ion tophoresis on osteoarthritis(OA) of rabbit knee joint in vivo.Methods The modified Hulth OA model was prepared with rabbits by anterior cruciate ligament desmotomy plus medial meniscus resection.Twenty-four model rabbits were randomly divided into three groups: Chinese medicine group: treated with Chinese medicine Shujin-Zhitong liquid by ion tophoresis for 20 minutes per day;positive control group: glucosamine hydrochloride was administered orally per day;blank control group: no treatment was administered to the model rabbits.Serum nitrogen monoxidum(NO) concentration was determined at the time points of prior model making,prior treatment,and 4 and 8 weeks of treatment.At 4 and 8 weeks of treatment,the apoptotic rate of chondrocytes was determined by flow cytometry,and the gene expressions of collagen Ⅱand aggrecan were determined by reverse transcription polymerase chain reaction(RT-PCR).Results The contents of serum NO in Chinese medicine group and positive control group were lower than that in blank control group at 4 weeks after treatment,and the content of serum NO in Chinese medicine group was lower than those in positive control group and blank control group at 8 weeks after treatment.The apoptotic rates in Chinese medicine group and positive control group were lower than that in blank control group at 4 and 8 weeks after treatment.Gene expression levels of collagen Ⅱ and aggrecan in Chinese medicine group and positive control group were higher than that in blank control group at 4 weeks after treatment,while the gene expression level in Chinese medicine group was higher than those in positive and blank control groups at 8 weeks after treatment.Conclusions The treatment of medicated permeation with traditional Chinese medicine Shujin-Zhitong liquid combined with ion tophoresis has shown good therapeutic effect on osteoarthritis(OA) of rabbit knee joint,the mechanism might be the inhibition of chondrocytes apoptosis and reduced secretion of inflammatory factors,and the improvement of cartilage repair by up-regulating related gene expression of chondrocyte.

Objective To investigate the expression of α-smooth muscle actin (α-SMA)in patients with valvular atrial fibrillation and study the relationship betweenα-SMA and atrial fibrosis in patients with valvular atrial fibrillation (AF).Methods For this study we enrolled 84 consecutive patients with rheumatic heart disease who were to receive cardiac surgery.The patients were divided into AF group (AF,n=39)and sinus rhythm group (SR, n=45).Their clinical data including baseline demographics,routine laboratory test and echocardiographics were collected before surgery.The right atrial tissue (0 .3-0 .5 cm3 )was disserted during the surgery.Right atrial fibrosis was observed by Masson staining.The mRNA expression ofα-SMA in atrial tissue were determined by Real-time quantitative PCR.Western blot was used to measure the protein expression ofα-SMA in atrial tissue.Results The two groups did not significantly differ in sex ratio,age,blood pressure,blood biochemical indicators or other aspects of medical history (P>0.05).However,left and right atrium diameters in AF group were significantly larger than those in SR group (P<0 .05 ).Masson staining suggested that collagen volume fraction and collagen content were significantly higher in AF group than in SR group (P<0 .05 ).The mRNA and protein expressions ofα-SMA in right atrial tissue were obviously higher in AF group than in SR group (coefficients P<0 .05 ).The mRNA and protein expressions ofα-SMA from right atrial tissue in the 84 patients were positively correlated with collagen content (coefficients of 0.587 and 0.607;P=0.029,0.014,respectively).Conclusion There is significant atrial fibrosis in patients with valvular atrial fibrillation,which is closely related to up-regulated expression ofα-SMA gene.

Background Immediately ocular rinsing is a key step for the prevention of eye tissue damage after acute chemical bums.A good ophthalmic rinsing solution can neutralize chemical substance and improve the prognosis of patients.Objective This study attempted to evaluate the therapeutic effect of self-made citric aciddisodium hydrogen phosphate buffer and citric acid-disodium hydrogen phosphate-potassium chloride buffer on corneal chemical burns in mice.Methods Citric acid-disodium hydrogen phosphate buffer solution (solution 1) and citric acid-disodium hydrogen phosphate-potassium chloride buffer solution (solution2) with the pH 7.4 were prepared.One hundred and twenty clean male C57 mice aged 6-8 weeks were randomized to two groups,and filter papers containing 1 mol/L H2SO4 or 0.15 mol/L NaOH were attached to the central corneas of the right eyes to create the acid or alkali burning models.Then the eyes were immediately rinsed by 40 ml solution 1,tap water or solution 2 according to the grouping and the model eyes without rinsing served as the control group.The corneal opacity was examined by slit lamp microscope and scored in 3,7 and 14 days after modeling.The percentages of corneal fluorescein staining,corneal neovascularization and corneal ulcer were analyzed.The study protocol was approved by Experimental Animal Ethic Commission of Second Military Medical University.Results In the corneal acid burning models,the number of eye with corneal opacity scored 1 in the solution 1 group,tap water group and solution 2 group was significantly more than that in the non-rinsing group in 3,7 and 14 days after modeling (all at P＜0.01);In 3 days after modeling,the numbers of eye scored 1 were more in the solution 1 group than those in the tap water group and solution 2 group (x2 =11.000,P =0.001;x2 =4.000,P =0.046).There were no differences in the eye number of different corneal opacity scores in 14 days after acid burning (all at P＞0.05).In 3,7 and 14 days after corneal alkali burning,the number of eyes with corneal opacity scored 1-2 was significantly increased in the solution 1 group,tap water group and solution 2 group compared with non-rinsing group (all at P＜0.01).The percentage of corneal ulcer in the solution 1 group,tap water group and solution 2 group was 7％,27％ and 13％,respectively,which was significantly lower than 73％ in the non-rinsing group (P =0.000,0.027,0.003),and no significant differences were seen in various time points after corneal alkali burning (all at P＞O.05).Corneal neovascularization occurred in 50％ mice in non-rinsing group in 14 days after acid burning.However,no neovascularization was seen in the mice of the solution 1 group,tap water group and solution 2 group in both acid and alkali burning mice.Conclusions Citric aciddisodium hydrogen phosphate buffer (pH 7.4) appears to be an effective emergency rinsing solution for corneal chemical burns and the rinsing solution with or without potassium chloride is not obviously affected to the prognosis of corneal chemical burns in the mice.

Objective:To investigate the expression of fibrillin-1 (FBN-1) in the atrium tissue of the patients with rheumatic heart valve disease complicated with atrial fibrillation(AF), and to explore its relationship with atrial fibrosis in the patients with valvular atrial fibrillation.Methods:Eighty-four consecutive patients with rheumatic heart valve disease underwent cardiac surgery were enrolled in this study.The patients were divided into AF group(n=39) and sinus rhythm group(SR group, n=45).The clinical data of patients were collected before operation.The right atrium tissue (0.3-0.5 mm3) was disserted during operation.The degrees of right atrial fibrosis of the patients in two groups were observed by Masson staining.Western blotting method was used to measure the protein expressions of FBN-1 in atrium tissue of the patients in two groups.Results:There were no significant differences in the gender ratio, age, blood pressure, blood biochemical indicators and other aspects of medical history between two groups(P>0.05);the diameters of left and right atrium of the patients in AF group were significantly larger than those in SR group(P<0.05).The Masson staining results showed that there was obvious fibrosis in AF group, and the collagen volume fraction and collagen level in AF group were significantly higher than those in SR group (P<0.05).The expression level of FBN-1 in right atrium tissue in AF group was obviously higher than that in SR group(P<0.05).The expression level of FBN-1 protein in right atrium tissue of the patients with valvular atrial fibrillation was positively correlated with the collagen level(r=0.544,P=0.021).Conclusion:There is obvious atrium fibrosis in the patients with valvular atrial fibrillation;it is closely related to the up-regulation of the expression of FBN-1 gene.

OBJECTIVE:To study the inhibitory effect of dihydrotanshinone(DTS)on human lung cancer GLC-82 cell and its mechanism. METHODS:After treated with 0(blank control),5,10,20,40,80 and 100 μg/ml DTS for 24 and 48 h,MTT as-say was used to measure the inhibition rates and IC50 of cells;cell apoptosis was detected by flow cytometry after treated with 17.85 μg/ml DTS for 12,24 and 48 h to calculate apoptotic rate;Western blot was used to detect the protein expressions of Bcl-2, Bax and Caspase-3. RESULTS:Compared with blank control group,different concentrations of DTS inhibited the proliferation of cells;24 and 48 h maximal inhibition rate were 54.48% and 64.95%,respectively;IC50 were 62.36 and 33.94 μg/ml. DTS could induce cell apoptosis in positive time dependent manner,and the range of inhibition rate was 5.6%-29.6%;Western blot showed DTS could down-regulate the expression of Bcl-2 protein and up-regulate the expression of Caspase-3 protein (P<0.01 or P<0.05). CONCLUSIONS:DTS have significant inhibitory effect on GLC-82 cells and also induce cell apoptosis,by a possible mech-anism of down-regulating the expression of Bcl-2 protein and up-regulating the expression of Caspase-3 protein.

Objective To analyze the operative therapeutic effect of proximal femoral nail antirotation blade (PFNA),dynamic hip screw (DHS) and femoral proximal locking compression plate (LCP)internal fixation in treatment with femoral intertrochanteric fracture of the aged.Methods Seventy-three aged patients with femoral intertrochanteric fracture were divided into 3 groups according to different internal fixation method:PFNA group (30 cases) was treated with PFNA internal fixation,DHS group (22 cases) was treated with DHS internal Fxation,LCP group (21 cases) was treated with LCP internal fixation.The operative time,intraoperative bleeding volume,fracture healing time,bad limb load time and hip joint function score were compared among the 3 groups.Results The operative time in PFNA group,DHS group and LCP group was (73.9 ± 8.5),(82.3 ± 10.6),(79.5 ± 11.2) min,there was statistical difference between PFNA group and DHS group (P ＜ 0.05),but there was no statistical difference between PFNA group and LCP group (P＞ 0.05).The intraoperative bleeding volume in PFNA group was significantly lower than that in DHS group and LCP group [(160.2 ± 15.6) ml vs.(208.4 ± 20.2) and (216.6 ml ± 24.9) ml],there was statistical difference (P ＜0.05).The fracture healing time and bad limb load time in PFNA group were significantly shorter than those in DHS group and LCP group [(97.4 ± 5.8) d vs.(101.2 ± 4.2),(110.0 ±8.3) d and (78.0 ±5.4) d vs.(85.9 ±7.6),(90.8±6.0) d],furtbermore DHS group were significantly shorter than those in LCP group,there were statistical differences among the 3 groups (P ＜ 0.05).There were no statistical differences in excellent and good rate of hip joint function score among the 3 groups (P ＞ 0.05).Conclusions PFNA is the ideal therapeutic method for femoral intertrochanteric fracture of the aged.It has better curative effect compared to the DHS and LCP,and has less surgical trauma and can shorten the recovery time.

AIM:To study the effects of extracellular potassium on the protein expression of wild-type HERG and its mutant L539fs/47.METHODS:Wild-type HERG (WT) or its mutant HERG-L539fs/47 (MT) were transfected into HEK293 cells for 36 h.The cells were incubated in different media containing 0.8, 4.3 or 10 mmol/L potassium.Af-ter 6 h of incubation, the protein expression of HERG was detected by flow cytometry.After 12 h of incubation, the locali-zation and quantity of the proteins were detected by laser confocal imaging and Western blot.RESULTS: Different from the retention of mutant protein in cytoplasm, wild-type HERG protein was mainly distributed in the cell membrane.The 2 proteins both increased with the changes of extracellular potassium.Flow cytometry showed that the fluorescence in the 2 groups both increased with the changes of extracellular potassium ( P<0.01 ) .The fluorescence in WT group was signifi-cantly higher than that in MT group (P<0.01).Western blot showed that mutant HERG protein included only one 60 kD band, different from the 135 kD and 155 kD bands in wild-type HERG, which were affected by the changes of extracellular potassium (P<0.05).CONCLUSION:The retention of HERG mutant L539fs/47 protein in the cytoplasm is more than wild-type HERG.Chronic high extracellular potassium keeps the stability of wild-type and mutant HERG proteins on the cell membrane.Chronic low potassium reduces the expression of HERG channel proteins in a time-dependent manner.

BACKGROUND: Lower extremity deep venous thrombosis is commonly seen after hip arthroplasty. The time of thrombosis after arthroplasty in senile patients with osteoporotic femoral neck fractures is little reported.OBJECTIVE: To investigate the time of perioperative lower extremity deep venous thrombosis in senile patients with osteoporotic femoral neck fractures undergoing hip arthroplasty. METHODS: The data of 178 patients with femoral neck fractures undergoing hip arthroplasty were analyzed, and were then allocated to anticoagulant and non-anticoagulant groups according to the preoperative use of low-molecular-weight heparins or not. Doppler ultrasonography of both lower extremity arteries was conducted at 1, 3, 5 and 5-7 days after injury, and 2, 5, 7 and 14 days postoperatively to monitor the lower extremity deep venous thrombosis, and the number and incidence of thrombosis in the two groups were recorded at different time points. The time of thrombosis occurred frequently after injury and surgery was determined through drawing the frequency distribution table and line chart.RESULTS AND CONCLUSION: (1) The incidence of thrombosis showed significant difference between anticoagulant (n=6, 8%) and non-anticoagulant (n=22, 21%)) groups (P < 0.05). (2) Thrombosis frequently occurred at 3 days after injury and 1 day postoperatively. (3) These results suggest that the thrombosis in patients with femoral neck fractures frequently occurs at 1-3 days after injury and 1 day after replacement. Additionally, active prevention measures after injury can significantly reduce the incidence of lower extremity venous thrombosis.

Objective To reveal the role of serum ACE2/Ang (1-7)in the occurrence of atrial fibrillation (AF)and find new targets for the prevention and treatment of AF by analyzing the correlation between the serum concentration of ACE2/Ang (1-7 )in patients with rheumatic valvular heart disease and the occurrence of AF. Methods We collected the basic clinical information and peripheral venous blood of patients with rheumatic heart valve disease (totally 46 patients,including 24 with AF and 22 with SR).ELISA method was used to detect the serum concentration of ACE2,Ang (1-7)and AngⅡ in the serum samples.Then the differences and correlation between the two groups were analyzed.Results In the AF group ① the diameter of the left atrium was significantly greater than that in the SR group [(60.70±3.08 vs.48.15±2.16)mm,P<0.05];② the serum concentration of AngⅡ was significantly higher than that in the SR group [(45.88±2.87 vs.35.78±1.08)pg/mL, P<0.05],AngⅡ and left atrium diameter were positively correlated (Pearson test,P<0.05);③ the serum concentrations of ACE2 [(7.87±0.74 vs.11.65±0.57)U/L,P<0.05]and Ang (1-7)[(146.05±17.61 vs. 321.71±36.50)pg/mL,P<0.05]were significantly lower than those in the SR group,and negatively correlated with left atrium diameter (Pearson test,P<0.05);④ the serum concentration of Ang (1-7)was negatively correlated with AngⅡ concentration (Pearson test,P<0.05).Conclusion For patients with rheumatic valvular heart disease,ACE2/Ang (1-7 )may play a protective role in the occurrence of AF via antagonizing AngⅡ and inhibiting atrial remodeling.