Objective To estimate the effect of leptin antibody on transforming growth factor (TGF)-β1 mRNA expression in hypertrophic scar model in rabbit ears.Methods Fifteen New Zealand white rabbits were included in this study.Three circular incisions which measured 7 mm in diameter and reached the perichondrium,were made in each ear of these rabbits to establish 90 models of hypertrophic scar.After the operation,these models were randomly and equally divided into 3 groups to be treated with topical sodium chloride physiological solution for 40 days (saline group),topical leptin antibody of 2 ng/ml for 40 days (leptin antibody group),and topical leptin antibody of 2 ng/ml for 14 days followed by injection of leptin antibody of 2 ng/ml once a week for 3 weeks (combination group).Scar tissue was resected from these rabbit ears at 40 days after the operation,followed by the determination of scar elevation index,histopathological examination by using haematoxylin and eosin staining,and quantification of TGF-β31 mRNA expression by real-time fluorescence-based PCR.SPSS 13.0 software was used for data processing.Statistical analysis was carried out by one-way analysis of variance.Results A significant decrease was observed in scar elevation index (2.33 ± 0.33 and 2.35 ± 0.22 vs.3.33 ± 0.41,both P ＜0.05) and TGF-β1 mRNA expression in the leptin antibody group and combination group compared with the control group,whereas no significant difference was observed between the leptin antibody group and combination group in either of the two parameters.Pathologically,there was an apparent proliferation of capillaries in the saline group with numerous irregularly and densely arranged fibroblasts with large nuclei,while relatively few fibroblasts with small nuclei,which were arranged in a more regular way,were observed in the leptin antibody group and combination group.Conclusion Leptin antibody treatment can reduce the expression of TGF-β1 mRNA in hypertrophic scar tissue in rabbit ears.

Objective To evaluate the value of high-frequency echocardiography in assessing Bax gene transfer influence on survival of heterotopic cardiac allograft in rats.Methods Thirty rat models of heterotopic heart transplantation were established.Group A received heart transplantation only; Group B received cyclosporin (CsA) after operatiom Group C received ultrasound targeted microbubble destruction (UTMD) combined with Bax-shRNA.All rats were tested by high-frequency echocardiography at day 1,3,6 after transplantation.The ultrasound parameters included left ventricular internal dimension diastole (LVIDd),left ventricular internal dimension systole(LVIDs),left ventricular ejection fraction(LVEF),left ventricular thickness (LVT),left ventricular thickening (LVTR) and so on.The pathologic examinations were carried out on five rats at day 6 after echocardiography exam.The other rats in each group were observed for the survival time of cardiac allograft.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by high-frequency echocardiography.②The survival time of allografts in group C was (16.21 ± 5.01)d,which was longer than that in group B [(11.14 ± 1.72)d,P ＜ 0.05] and group A [(7.26 ± 1.50)d,P ＜0.01].③LVT index got higher after transfection.At day 6,LVEF of group A was lower than group B and C markedly(P ＜0.05) while no significant difference was found between group B and C(P ＞0.05).At day 3 and 6,LVT of group A and B were higher than group C (P ＜0.05) while LVTR was lower(P ＜0.05).Conclusions The Bax gene transfer influence on survival of heterotopic cardiac allograft in rats could be evaluated accurately by high-frequency echocardiography which could assess cardiac structure and function.LVT and LVTR can be considered as early evaluation indexes for its high sensitivity over LVEF.

Objective To investigate the clinical and epidemiologieal feat ures of epidemic cerebrospinal meningitis in recent 3 years,aiming to develop the strategies for controlling this disease.Methods A retrospective analysis was conducted in 65 hospitalized patients with epidemic cerehrospinal meningitis from 2003 to 2006.Chi-square test was used for statistical analysis.Results The majority of these 65 patients were juvenile and adult,accounting for 44.6%(29 cases)and 35.4%(23 cases),respectively,while the infant patients account for the smallest percentage,only 6.2%(4 cases).Most cases occurred in spring(from February to April).The clinical features of most patients belonged to common type(72.3%),followed by fulminant type(27.7%).Five cases(7.7%)died,all of whom were fulminant cases.Totally 33 strains of Neisseria meningitidis were isolated,with positive culture rate of 50.8%(33/65 cases).The positive rate of blood culture was 44.6%(29/65 cases)and that of cerebrospinal fluid cuhure was 31.4%(16/51 cases).Isolates of Neisseria meningitidis were still highly sensitive to penicillin,ceftriaxone and cefotaxime.However,the resistance rates of strains to compound sulfamethoxazole,gentamycin and ciprofloxacin were all above 80%.Resuhs of serological typing revealed thai 82.8%(53/64 cases)cases belonged to group C.There were more severe cases and higher death rate in patients infected by group C meningococcus.Conclusions Serogroup C of the meningococcus have become the preponderant strains in Anhui Province.Most of the patients infected hy group C are J uveniles and adults.Penicillin and the third generation cephalosporins are still highly aclive againsl Neisseria meningitidis.

Objective To evaluate the diagnostic value of high frequency echocardiography on mice with doxorubicin-induced cardiotoxicity and find out the most valuable monitoring indexes for early doxorubicin-induced myocardial damage.Methods Forty mice were divided into doxorubicin-injection group (ADM group) and control group randomly.Twenty mice in each group were injected with doxorubicin and saline at the same dose for 5 weeks continuously.The parameters of cardic function of ADM group and contol group were observed by high-frequency echocardiography after infection.Brain natriuretic peptide (BNP) tested by euzymelinked immunosorbent assay(ELISA) and pathological findings were used as the golden standard.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by transthoracic high-frequency echocardiography.②After 5 weeks intervention,weight,heart rate,left ventricular ejection fraction (LVEF),left ventricular fractional shortening (LVFS),E/A,E wave deceleration time(EDT) and other parameters showed marked difference between ADM group and control group(P ＜0.05).③The decrease of diastolic function was earlier than systolic function in ADM group.④LVEF and E/A presented good negative linear correlation to BNP(r =-0.622,P ＜0.05; r =-0.593,P ＜0.01).EDT showed positive linear correlation to BNP(r =0.501,P ＜0.05).Conclusions Cardic function of mice with doxorubicin-induced myocardial damage could be early evaluated accurately by high-frequency echocardiography,which contained good relativity with BNP.

Objective To establish a simple and efficient method for developing a keloid model in nude mice with human keloid-derived fibroblasts.Methods Twenty-seven female BALB/c nude mice were randomly divided into five groups with 5,5,5,8 and 4 mice in group A,B,C,D and E respectively.The mice in group A,B and C were inoculated with 0.1 ml of suspension containing human keloid-derived fibroblasts at concentrations of 1.0 × 104,3.0 × 104 and 5.0 × 104 per microliter Matrigel,respectively,at the right axillary fossa.The tumors that formed in one mouse in group C were taken out,and cut into several parts measuring 5 mm × 5 mm × 5 mm in size,which were then subcutaneously transplanted into the right axillary fossa of mice in group D.The mice in group E were subcutaneously injected with 100 μl of Matrigel and served as the control group.The formation of tumor in mice was observed by naked eyes,and the size of tumors was measured until day 30 after tumor formation in group A,B and C as well as after tumor transplantation in group D.Mice were sacrificed on day 30 after tumor formation,and histopathologic examination was performed to analyze histological features of transplanted tumors and pathological changes in visceral organs such as heart,liver,spleen,lung and kidney.Results The tumor formation rate was consistently 100％ in group A,B and C,and the time required for tumor formation was (90.20 ± 3.96),(61.00 ± 2.92) and (39.60 ± 3.20) days in group A,B and C respectively.There was a significant difference in tumor volume on the 30th day after tumor formation between group A,B and C ((288.34 ± 25.29) vs.(1 370.63 ± 105.24) vs.(1 940.98 ± 184.37) mm3,F =138.74,P ＜ 0.05).The size of implanted tumor mass in group D firstly increased,then gradually decreased,but began to continuously increase since the 14~ day,and tumor finally formed in 7 out of 8 mice.There was no evidence of tumor formation in group E.Histopathologic examination showed uniform histological manifestations,which were similar to those of human scar,in tumor tissues from mice in group A,B,C and D.Neither pathological changes nor metastases were observed in visceral organs of these mice.Conclusion Keloid-bearing nude mouse model can be established by subcutaneous inoculation with human keloidderived fibroblasts,or by subcutaneous transplantation of tumor masses of a certain size that have formed in nude mice.

Objective: To explore the effects of N-(4-hydroxyphenyl) retinamide (4HPR), 4HPR liposome (4HPR-L), and 4HPR lipid microbubble (4HPR-LM) combined with ultrasound on proliferation, apoptosis, and cell cycle of human keloid fibroblasts (Fbs). Methods: (1) 4HPR-L and 4HPR-LM were prepared by hydration ultrasonic method. The appearance morphology, particle size distribution, Zeta potential, loading drug concentration, encapsulation efficiency, and drug loading rate of 4HPR-L were investigated by high performance liquid chromatography, dynamic light scattering, and transmission electron microscope. (2) Human keloid Fbs were cultured and divided into 13 groups by random number table (the same grouping method below), with 6 wells in each group. Cells in control group were given no treatment, while cells in 12 ultrasound groups including 0.5 W 30 s group, 0.5 W 60 s group, 0.5 W 120 s group, 0.7 W 30 s group, 0.7 W 60 s group, 0.7 W 120 s group, 1.0 W 30 s group, 1.0 W 60 s group, 1.0 W 120 s group, 1.5 W 30 s group, 1.5 W 60 s group, and 1.5 W 120 s group were treated by ultrasound with corresponding parameters. The cells viability was measured by a microplate reader after 24 hours of routine culture. Another batch of human keloid Fbs were divided into 5 groups, with 6 wells in each group. Cells in control group were given no treatment, while cells in 1, 10, 20, and 50 μg/mL blank lipid microbubble groups were treated with blank lipid microbubbles in corresponding mass concentration. The cells viability was measured as before after 24 hours of routine culture. Another batch of human keloid Fbs were divided into 6 groups, with 12 wells in each group. Cells in control group were given no treatment, while cells in 1, 10, 20, 50, and 100 μg/mL 4HPR-L groups were added with 4HPR-L carrying corresponding mass concentration of 4HPR. The cells viability in 6 wells of each group was detected after 24 and 48 hours of routine culture, respectively. Another batch of human keloid Fbs were divided into 4 groups, with 6 wells in each group. Cells in control group were given no treatment, while cells in 4HPR, 4HPR-L, and 4HPR-LM+ ultrasound groups were treated with 4HPR, 4HPR-L, and 4HPR-LM (all the mass concentration of 4HPR was 20 μg/mL), respectively, and cells in 4HPR-LM+ ultrasound group were given 0.5 W 60 s ultrasound treatment immediately after drug administration. The cells viability was measured as before after 24 hours of routine culture. (3) Another batch of human keloid Fbs were divided into control group, 4HPR group, 4HPR-L group and 4HPR-LM+ ultrasound group, with 3 wells in each group, and the cells in each group were treated as before. Apoptosis of the cells was detected by flow cytometer after 24 hours of routine culture. (4) Another batch of human keloid Fbs were grouped and treated as in (3), and then the cell cycle distribution was detected by flow cytometer after 24 hours of routine culture. Data were processed with one-way analysis of variance and t test. Results: (1) 4HPR-L particles had a spherical or spheroidal structure and were uniform in size, with particle size of (100.1±1.3) nm and Zeta potential of (-34.3±2.3) mV. The mass concentration of 4HPR in 4HPR-L solution was about 1 400 μg/mL, with the encapsulation efficiency of (95.8±1.2)% and drug loading rate of (8.3±0.4)%. (2) The viability of cells in the 12 ultrasound groups was higher than 93.0%, and the viability of cells in 1, 10, 20, and 50 μg/mL blank lipid microbubble groups was higher than 95.0%. The viability of cells in 1 μg/mL 4HPR-L group at administration hour 24 was similar to that at 48 (t=0.393, P>0.05). The viability of cells in 10, 20, 50, and 100 μg/mL 4HPR-L groups at administration hour 24 was significantly higher than that at administration hour 48 (t=44.593, 22.961, 32.224, 35.337, P<0.01). The viability of cells in 4HPR group, 4HPR-L group, and 4HPR-LM+ ultrasound group was (47.3±0.7)%, (42.3±1.7)%, and (38.6±0.8)%, respectively. The viability of cells in 4HPR group was significantly higher than that in 4HPR-L group and 4HPR-LM+ ultrasound group (t=4.551, 15.895, P<0.05 or P<0.01). The viability of cells in 4HPR-L group was significantly higher than that in 4HPR-LM+ ultrasound group (t=-3.360, P<0.05). (3) The percentages of total apoptotic cells in 4HPR group, 4HPR-L group, and 4HPR-LM+ ultrasound group were (32.8±2.4)%, (42.5±2.4)%, and (58.5±6.3)%, respectively, which were significantly higher than the percentage of control group [(14.9±1.6)%, t=8.748, 13.637, 9.500, P<0.01]. The percentages of total apoptotic cells in 4HPR-L group and 4HPR-LM+ ultrasound group were significantly higher than the percentage in 4HPR group (t=4.049, 5.393, P<0.05 or P<0.01), and the percentage of total apoptotic cells in 4HPR-LM+ ultrasound group was significantly higher than that in 4HPR-L group (t=3.371, P<0.01). (4) The percentage of G2/M phase cells in 4HPR group was higher than that in control group, but there was no statistically significant difference (t=2.107, P>0.05). The percentage of G2/M phase cells in 4HPR-L group was significantly higher than that in 4HPR group or control group (t=18.169, 30.026, P<0.01). The percentage of G2/M phase cells in 4HPR-LM+ ultrasound group was significantly higher than that in 4HPR-L group, 4HPR group, and control group (t=4.932, 25.854, 66.231, P<0.01). Conclusions 4HPR can inhibit proliferation, induce apoptosis, and arrest G2/M phase of human keloid Fbs, and the effects of 4HPR-LM combined with ultrasound are better than those of 4HPR-L and free 4HPR.

Objective To silence Bax gene in H9c2 cells by ultrasound-targeted microbubble destruction(UTMD) combined with polyethylenimine.Methods The shRNA expression plasmid targeting Bax gene was co-treated with microbubbles and polyethylenimine.Then the mixture added to H9c2 cells were irradiated by ultrasound.Grouping:① plasmid group;② Lipofectamine2000 + plasmid group; ③ UTMD + plasmid group;④ PEI + UTMD + plasmid group;⑤ PEI + SonoVue + plasmid group (without ultrasound irradiation).The transfection efficiency was assessed by fluorescence microscope and FCM.The expression of Bax mRNA and protein were detected by RT-PCR and Western Blot.Results Bax shRNA wasn't destroyed by ultrasound exposure.The transfection efficiency of PEI + UTMD + plasmid group was higher than any other groups obviously (P ＜0.05).The result of RT-PCR and Western Blot showed inhibition efficiency of Bax mRNA and protein expression in PEI + UTMD + plasmid group were (69.41 ± 4.91) ％ and (64.09 ± 2.38)％ separately,which were higher than any other groups significantly (P ＜0.05).Conclusions UTMD combined with polyethylenimine can inhibit the expression of Bax gene in H9c2 cells effectively.

Objective To explore the effect of repaglinide intensive treatment on islet β-cell function and long-term control of blood glucose in newly diagnosed type 2 diabetic patients. Methods Self-control and inter-group control prospective study was conducted in 80 newly diagnosed type 2 diabetic patients who were treated with short-term repaglinide intensive treatment and islet β-cell function was assessed by 75 g oral glucose tolerance test (OGTT) before and after repaglinide treatment. The changes of △I30/△G30 ratio, blood lipid, HOMA A and HOMA B were examined. Results After treatment, in successful group, middle group and defeat group, the fasting plasma glucose levels were decreased from 8.9±1.5, 8.6±1.6,9.0±2.0 to 5.0±1.4,6.3±0. 7,6.5±0. 9 mmol/L, 0. 5 h postprandial glucose levels were decreased from (12.6±1.6, 12.6±1.5, 12.4±1.3 to 8.4±1.0, 6.8±0. 7, 8. 6±0. 9)mmol/L,and 2 h postprandial glucose levels were decreased from (13.0±1.2, 13. 1±1.3, 13. 3±1.4 to 9.2±0.9, 6.6±0. 7, 9.2±0. 9)mmol/L,respectively (all P <0. 005). The ratio of △I30/△G30 was increased froml. 69±0. 31, 1.72±0. 33, 1.79±0. 36 to 4. 47±0. 62, 4. 42±0.46,12. 00±0.46 in the three groups, respectively (P<0.05). HOMA B was significantly improved (P<0. 05), while triglycerides and HOMA A were decreased(P<0. 05). The levels of fasting blood glucose and postprandial blood glucose in 21 patients were maintained within normal range for more than six months. There were significant differences in the ratio of △I30/△G30, age, repaglinide dosage and the time of reaching target of glucose [4.47±0.62 vs. 2. 0± 0.46; 39±8 vs. 56±9; 2.0±1.5 vs. 5.0±2.5; 32.4±8.0 vs. 53.3±7.6; all P<0.05] between successful group and defeat group. Conclusions The short-term intensive treatment with repaglinide can significantly improve the early secretion phase of insulin and the islet β-cell function, reconstruct of the physiological model of insulin secretion and relieve the disease.

Objective To primarily study the possible biological significance of yon Willebrand factor (vWF) expression at the fracture site in the mechanism of fracture healing. Methods A total of 28 male SD rats were selected to set up femoral fracture models. Then, tissue samples from fracture site were randomly taken at days 2, 3, 5, 7, 10, 14 and 21 after establishment of the models, with 4 rats at each time point, vWF expression at fracture site and the relationship of vWF expression with osteoblasts and chondroblasts were observed by using computer image analysis system. The biological significance of vWF expression at fracture site in mechanism of fracture healing were analysed based on the results of vWF expression of in vitro cultured rat osteoblasts. Results The expression of vWF at every time point was positive and distributed disproportionally irregular at bone trabecula and soft tissue of the fracture site. vWF expressed positively in the extracellular matrix around the chondroblasts. Conclusions During the course of fracture healing, there may be vWF-platelet activation pathway that can regulate differentiation and proliferation of local osteoblasts and chondrocytes and activate platelets secreting growth factors such as PDGF, IGF-1, EGF and TGF-a, as creates a sound microenvironment for fracture healing.

This study aims to investigate the change of plasma concentration of digoxin (DIG) in rats with ovariectomy. Twelve female SD rats were randomly assigned into ovariectomized group and sham group (n = 6). All rats plasma was collected after a single dose of 2 mg x kg(-1) DIG administrated orally, serum DIG concentration was determined by LC-MS/MS. The level of P-gp in the intestinal was analyzed by Western blotting. Pharmacokinetic calculations were performed on each individual using DAS 2.0 practical pharmacokinetic software. Compared with the sham group, C(max) of ovariectomized group decreased significantly (P < 0.01). There was no significant difference of AUC(0-t), and the level of P-gp was elevated in ovariectomized group. It was found that C(max) of DIG was significantly reduced after ovariectomy, and the change was associated with the decreased level of estrogen, which contributes to the increased level of P-gp.