Objective To establish and verify the animal model of sclerotic skin induced by bleomycin(Blm).Methods To establish a mouse model for scleroderma in C3H/He mice by repeated local injection of100?L of Blm(200?g/mL)everyday for3weeks.Then,the specimens of skin,lung and serum were examined.Results After3week local Blm injections,an intense dermal sclerosis was shown in C3H/He mice.Compared with the control skin,increased dermal thickness and increased collagen histo-chemical index were found(P

Objectives To study the effects of traditional Chinese herbs, Wen Yang Huo Xue Decoction (herbs of removing blood-stasis and warming the kidney-yang) and Salvia miltiorrhiza, on the animal model of sclerotic skin. Methods A mouse model for sclerotic skin was established in C3H/He mice by repeated local injections of bleomycin for 3 weeks. Wen Yang Huo Xue Decoction was given orally, and Salvia miltiorrhiza orally or intravenously, to the mouse model. The administrations started either simultaneously or after 3 weeks' injections of bleomycin. Mouse skins and lungs were examined histopathogically, and sera were tested for autoantibodies. Results The administrations of herbs, started either at the beginning or at the time sclerosis was induced, caused no significant alleviation of dermal sclerosis by the end of 5 weeks' treatment. After 8 weeks' administrations of herbs, the dermal thickness reduced and collagen histochemical index decreased significantly, especially in group Wen Yang Huo Xue Decoction was given orally and in group Salvia miltiorrhiza was given intravenously at early stage (P

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Objective:To determine ochratoxin A ( OTA ) in Chinese herbs by high performance liquid chromatography-tandem mass spectrometry method ( HPLC-MS/MS) . Methods: The samples were extracted by 80% methanol water solution and purified by immunoaffinity column. The chromatographic separation was carried out on a C18 column, the mobile phase was methanol-acetonitrile-0. 01 mol·L-1 ammonium acetate, and then OTA was detected by MS/MS in an ESI(-)-MRM mode. Results:The limit of detection was 0. 1 μg·kg-1 , the average recoveries ranged from 84. 8% to 91. 2%, and the relative standard deviations ( RSD) ranged from 3. 6% to 8. 1%(n=9). Conclusion:The method is accurate,sensitive and simple, and suitable for the determination of ochratoxin A in Chinese herbs.

Objective To investigate the clinicopathological characteristics,diagnosis and therapy,as well as the prognosis of accessory breast cancer. Methods Twenty-two cases were diagnosed as accessory breast cancer from Jan 1,1984 to Dec 31,2008, their clinicopatholgical data were analyzed retrospectively.Results Up to Mar. 1, 2010,2 cases had local recurrence;7 cases had long-distance metastasis;6 cases died. In the current study,the 5-year survival rate of accessory breast cancer was 43. 7%. Conclusions Accessory breast cancer is aggressive. The diagnosis was mainly based on clinical characteristics and postoperative pathology. The combined therapies may improve the survival rate.

ObjectiveTo study transcriptional characteristics of type Ⅲ procollagen gene in systemic scleroderma (SS)-derived fibroblast clones and their regulation by Radix Salviae miltiorrhizae(RSM).Methods Eight fibroblast clones with different collagen-producing capacity were previously obtained from patients with SS and normal human controls.Recombinant plasmids containing different deletions of the human alpha 1 chain of type 3 procollagen(COL3A1) gene promoter were constructed,and transiently transfected into the fibroblast clones.Dual-luciferase reporter assay system was used to evaluate the activities of these recombinants in the fibroblast clones and to select a proximal transcriptional regulatory sequence.Then,the fibroblast clones were transfected with the plasmid containing the selected regulatory sequence(phCOLH30.1) followed by the treatment with RSM injection(1 g/L) and active monomers of RSM,including salvianolic acid B(5 mg/L),tanshinone Ⅱ A (5 mg/L),danshensu(20 mg/L) and protocatechuic aldehyde(5 mg/L),for 48 hours.The transfected fibroblast clones receiving no drug treatment served as the water-soluble control,and those treated with only dimethyl sulfoxide as the lipid-soluble control.Subsequently,the fibroblasts were lysed and subjected to the quantification of cellular proteins and determination of luciferase activity.The activity of recombinant promoters was compared by t test for the selection of proximal transcriptional regulatory sequence,and the activity of phCOLH30.1 by two-way analysis of variance in the RSM-interfering test(if there was interaction,one-way analysis of variance was conducted; and if there was no interaction,the main effect was tested after the removal of interaction item).ResultsOf the 6 recombinants,the recombinant containing COL3A1 proximal promoter from -96 bp to +16 bp(phCOLH30.1) showed the highest transcriptional activity in nearly all of the fibroblast clones,and the activity was positively correlated with the collagen-producing capacity of fibroblast clones.Compared with the water-soluble control,RSM injection significantly downregulated the activity of phCOLH30.1 in fibroblast clones with high and low collagen-producing capacity from patients with SS (2.261 ± 0.619 vs.3.879 ± 0.309,1.462 ± 0.291 vs.2.150 ± 0.262,both P ＜ 0.01) and normal human controls (1.681 ± 0.263 vs.3.039 ± 0.271,1.121 ± 0.361 vs.2.223 ± 0.247,both P ＜ 0.01),salvianolic acid B decreased the phCOLH30.1 activity in SS-derived high collagen-producing fibroblast clones (2.309 ± 0.524,P ＜ 0.01 ) and in the normal control fibroblast clones with high and low collagen-producing capacity (2.126 ± 0.320 and 1.976 ± 0.362,both P ＜ 0.05).Tanshinone Ⅱ A only downregulated the phCOLH30.1 activity in SS-derived high collagen-producing fibroblast clones compared with the lipid-soluble control(2.975 ± 0.666 vs 5.379 ± 0.238,P ＜ 0.01 ).Neither danshensu nor protocatechuic aldehyde showed inhibitory effects on phCOLH30.1 activity in SS-derived or normal control fibroblast clones.ConclusionsThe type Ⅲ procollagen gene is activated at the transcriptional level in high collagen-producing fibroblast clones from patients with SS,and the activation could be suppressed by RSM injection,salvianolic acid B and tanshinone Ⅱ A.

Objective To present the surgical results of severe scoliosis treated with one stage anterior release and posterior instrumentation. Methods 36 patients of severe scoliosis received one stage anterior release and posterior correction during July 1997 to January 2003. There were 9 males and 27 females with an average age of 17.2 years, including 33 idiopathic scoliosis and 3 neurofibromatosis scoliosis. The Cobb's angles of scoliosis were from 85? to 116? with a mean of 96.2?. 18 cases were found with single thoracic curve, 7 cases double thoracic and lumbar curves, 6 cases thoracolumbar curve, 4 cases lumbar curve, and 1 case double thoracic and lumbar curves with associated lumbar curve. 20 cases had abnormal sagittal profile. Three-dimensional instrumentations such as CD (4 cases), CD-Horizon (5 cases), TSRH (10 cases) or Isola (17 cases) were used in posterior procedure after anterior release under the same anesthesia. 31 cases of this group received thoracoplasty. Results The coronal correction was achieved 48.5% on average. The sagittal profile of the spine was distinctly ameliorated. The sagittal balance was restored or maintained in 80.6% of the patients. There were no severe neurological complications, hook displacement, rod breaking and deep infection at follow-up. One case of traumatic pleurisy occurred after surgery and one pseudarthrosis at 2 years later. One patient demonstrated imbalance 11 months after surgery. Two patients presented loss of correction more than 10? at one year follow-up (5.2? on average). Conclusion The result of one stage posterior correction associated with anterior release in treatment of severe scoliosis is satisfactory. Appropriate selection of cases, detailed assessment, SEP monitoring and wake-up test during surgery are helpful to reduce severe complications. The long term results need to be followed-up.

Objective To find out if different doses of melatonin have different roles in the cause or treatment of idiopathic scoliosis, and to illuminate whether melatonin can be a substitution in the treatment of pinealectomized chickens. Methods Fifty Little-Shaoxing chickens, in which pinealectomy was performed when 3-day-old, were divided into five groups. Ten served as control group, were kept in light-dark (12h: 12h) cycle, 500 lx in daytime and 0-5 lx in nighttime after surgery. Ten served as M-5 group, were given 5 mg/kg (5 milligrams per kilogram of body weight) melatonin twice every day. Ten as M-10 group, given 10 mg/kg melatonin abdominally twice every day, ten as M-20 group, given 20 mg/kg melatonin twice every day, and ten as M-40 group, given 40 mg/kg melatonin twice every day. The light-dark (12h: 12h) cycle was the same as control group. Radiological examinations were performed on all chicken spines for scoliosis when at 3 months old. Measured of the angle of the scoliosis was taken by Cobb's technique, and the data was analyzed by statistical software. Results Overall, there was 25 chickens developed to scoliosis in all when the chickens were 3 months. The attack rate was only 56%; this rate was consistent throughout all experimental groups. In the control group, 6 chickens had obvious scoliosis, and 5 in M-5 group, 5 in M-10 group, 4 in M-20 group, and 5 in M-40 Group. And the average Cobb's angle of control group was 26.7?. There were 23.5?, 21.7?, 24.5? and 23.2? in M-5, M-10, M-20 and M-40 respectively. There was no statistical significance in the average Cobb's angle and the incidence of scoliosis among all groups. It was determined that neither the prevalence nor the pattern of the scoliosis was affected by the therapy in any of the experimental groups. Conclusion Different dose of melatonin can't reduce the rate of scoliosis in the pinealectomized chickens, and also can't prevent the development of the experimental scoliosis. The results of this study raise doubts regarding the role of melatonin in the development of scoliosis after pinealectomy in the young chicken. Melatonin acting as a vicarious therapeutic tool in the treatment of idiopathic scoliosis is premature, deserved further research.

Objective To explore the correlation between heart rate variability ( HRV ) and multiple system atrophy( MSA) .Methods HRV was performed in 34 MSA patients and 32 healthy controls.The parameters of HRV study was compared, parameters studied included time domain and frequency domain.Single factor analysis andreceiver operating characteristic ( ROC ) curves were used to analyze the detection of parameters of HRV. ResultsThe HRV of MSA showed significant decreased from the health controls: root mean square successive difference of intervals (rMSSD) [(51.6 ±29.4)ms vs (70.1 ±34.1)ms,P=0.023],percentage of interval differences of successive N-N intervals greater than 50 ms(pNN50%) [ 4.2% (1.8%,9.5%) vs 9.6%(5.1%, 16.2%),P=0.005], coefficient of variation(CV) [(0.06 ±0.01) vs(0.08 ±0.02),P=0.005],very low frequency(VLF) [1003.4(586.5,1702.9)ms2vs1825.5(1407.2,2670.1)ms2,P=0.000],lowfrequency(LF) [162.9(90.6,337.4) ms2 vs 375.5(210.4,559.5) ms2,P=0.001], high frequency(HF) [164.9(77.5,470.7) ms2 vs 349.1(209.7,738.5)ms2,P=0.005].The areas under ROC in patients with MSA for VLF was 0.703(S=0.053), while the areas for CV, LF, HF and pNN50%were 0.667(S=0.054), 0.660 (S=0.055), 0.650( S=0.055) and 0.640( S=0.055) ,the results all have statistical significance(all P<0.05).The cutoff value of VLF for diagnosing MSA in the early stage was 1240.85, the sensitivity and specificity were 86.7 and 55.4(95%CI:0.599-0.807,P=0.002) .Conclusion MSA has sympathetic and vagus nerve involvement at the early stage disease, resulting dysfunction of cardiovascular autonomic system,VLF may have high drgree of assessing accuracy of its dysfunction.

Objective To study the transcriptional regulation of type Ⅰ procollagen gene in systemic scleroderma(SS)-derived high collagen-producing fibroblast clones by Radix Salviae Miltiorrhizae(RSM).Methods Fibroblast clones with different collagen-producing capacity were previously obtained from patients with SS and normal human controls,and divided into 5 groups to be treated with RSM(1 g/L)injection,its water-soluble active monomers including sodium danshensu(20 mg/L),salvianolic acid B(5 mg/L)and protocatechuic aldehyde(5 mg/L),and lipid-soluble active monomer(tanshinone Ⅱ A,5mg/L)respectively.The fibroblast clones incubated with no drugs served as the water soluble negative control group,and those with dimethyl sulfoxide(DMSO)as the lipid soluble negative control group.MTT assay was performed to evaluate the proliferation of the fibroblast clones after 1-,3-,5-,and 7-day treatment,transient transfection and dualluciferase reporter assay system to quantify the relative activity of collagen type Ⅰ,alpha 1(COL1A1)proximal promoter in these fibroblast clones.Results The inhibitory effect of RSM and its active monomers on the proliferation of fibroblast clones was inapparent within the initial 3 days(P ＞ 0.05),but was enhanced with incubation time.A significant difference was observed in the proliferation level of fibroblast clones between RSM group and water-soluble negative control group on day 5(q′ =3.22,P ＜ 0.01),between RSM,salvianolic acid B,protocatechuic aldehyde groups and the water-soluble negative control group(q′ =4.74,3.03,2.56,all P ＜0.05)on day 7,and between tanshinone Ⅱ A and lipid-soluble negative control group on day 5 and 7(t =2.22,2.15,both P ＜ 0.05).RSM injection,tanshinone Ⅱ A and protocatechuic aldehyde significantly inhibited COL1A1 proximal promoter activity in SS-derived and normal control fibroblast clones(all P ＜ 0.01),and the former two drugs preferentially downregulated COL1A1 proximal promoter activity in SS-derived high collagenproducing fibroblast clones.Significantly different COL1A1 proximal promoter activity was observed in SS-derived high and low collagen-producing fibroblast clones between water-soluble negative control group and RSM injection group(12.019 ± 0.830 vs.4.445 ± 1.061,5.388 ± 0.480 vs.2.856 ± 0.597,F=31.78,P＜ 0.01),and between lipid-soluable negative control group and tanshinone Ⅱ A group(14.155 ± 0.672 vs.9.638 ±0.854,4.299 ± 0.252 vs.3.192 ± 0.450,F=24.10,P＜ 0.01).Conclusions RSM inhibits the transcription of COL1A1 gene in SS-derived high collagen-producing fibroblast clones,which may be mainly attributed to tanshinone Ⅱ A and protocatechuic aldehyde.