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 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.

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Adult ; Causality ; Cross-Sectional Studies ; Efficiency ; Female ; Humans ; Male ; Middle Aged ; Occupations ; Social Support ; Stress, Psychological ; Surveys and Questionnaires

This study was aimed to compare the activation of daidzein and puerarin on antioxidation, NO and NOS suppression in vitro . The RAW264 . 7 cell line was used to prepare radical reaction model induced by LPS (1 μg/mL). MTT method was adopted to detect cytotoxicity of daidzein and puerarin. The DCFH-DA probe and confocal microscopy were used to examine the antioxidant ability of daidzein and puerarin. Griess reagent was adopted to test the NO level in the culture medium. And chemical colorimetry was used to detect the content in RAW264.6 cells. The results showed that daidzein and puerarin can significantly suppress the NO and T-NOS expression in RAW264.7 cells induced by LPS. It was concluded that there was no difference on the activation of antioxidant free radical between daidzein and puerarin . In this regard , daidzein can be the used as substitutes of puerarin .

Abstract: Fifteen novel ligustrazine-tetrahydroisoquinoline derivatives were designed and synthesized according to the association principle of pharmaceutical chemistry. The structures were identified by IR, NMR and ESI-MS. The inhibitory activities of platelet aggregation induced by ADP and AA have been measured by Bron method. Preliminary pharmacological results showed that compounds 7g, 7h and 7n had potent inhibitory activity against platelet aggregation induced by AA, and the compound 7o showed significant inhibitory activity against platelet aggregation induced by ADP.
Drug Design ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; chemical synthesis ; chemistry ; Pyrazines ; chemical synthesis ; chemistry ; Tetrahydroisoquinolines ; chemical synthesis ; chemistry

Objective To analyze the clinical features of patients with subclinical pheochromocytoma(PHEO).Methods Review of clinical features of 22 patients with subclinical PHEO treated in PUMC hospital from 1997 to 2007.Results All patients were asymptomatic.24hr-urinary catecholamine excretion was detected normal in 10 of 22 cases,while increased in the others.Sixteen patients were prepared with ?-receptor blocker before operation.During the operation,BPmax(maximal blood pressure) before tumor resection,BPmin(minimal blood pressure) after resection and ?BP(BPmax-BPmin) were(163?34)/(86?20)mmHg,(105?12)/(61?10)mmHg and(58?37)/(25?21)mmHg,Respectively,in the prepared group.They were(169?36)/(104?20)mmHg,(97?18)/(56?13)mmHg and(71?48)/(48?29)mmHg in the other 6 cases without ?-blocker preparation.DBPmax and ?DBP in the prepared group were significantly lower than the unprepared group.Conclusion Most patients with subclinical PHEO have increased catecholamine secretion.Blood pressure is fluctuant greatly during operation in some patients.Patients should be treated with ?-receptor blocker preoperatively in order to decreasethe operation risk.

Objective To investigate the immunologic classification in the patients with acute leukemia (AL) in Xinjiang of China. Methods A panel of monoclonal antibodies (MOAb) and indirect immunofluorescence assay by fluoromicroscope was used to determine the pretherapy immunophenotype of 450 AL. Results 106 cases of acute lymphoblastic leukemia (ALL), 334 cases of acute myelogenous leukemia (AML), and 10 cases belonged to FAB unclassified acute leukemia (UAL) were unalysed. The expression of myeloid antigens in of ALL was seen in 15 % of 106 cases, and lymphoid-associated antigens were expressed in 25 % of 334 AML cases. The most frequently expressed antigen was CD7. The expression of myeloperoxidase (MPO) gene in 295 cases of AL were studied. The expression of MPO gene was observed in positive one of 81 ALL cases, and myeloid cells had different expression for MPO gene. Of the 9 cases of UAL, 6 cases were positive for MPO gene. There were no statistic differences of the expressions of the ALL stages between Han and Wei nationality. The order of myeloid markers expression in AML was as follows: CD_(33)>CD_(13)>CD_(15) inthe Han nationality, and the order of myeloid markers expression in AML was displayed CD(15)>CD(33)>CD_(14) in Wei nationality. Conclusion Analysis of immunophenotype assured accurate lineage diagnosis of AL. Combinatively analyzing the characteristics of AL on morphology, cytochemistry, immunology and MPO mRNA expressions were significant to the diagnosis and therapy of AL.

ObjectiveTo diagnose and classify 249 patients with myelodysplastic syndrome (MDS) according to the WHO standards.MethodsAccording to the WHO standards,cell morphology,cytogenetics,immune phenotype and bone marrow pathologic biopsy in 249 cases of MDS were analyzed.ResultsGreat shape and oval cell of mature erythrocyte could be observed in all MDS patients peripheral blood. The incidence of immature erythrocyte,immature granulocyte,pelger-like abnormal nucleus and neutrophils cells without granular increased with subtypes progressing.These abnormal characteristics and proportion tended to more apparent with MDS subtypes progressing.With the dynamic follow-up,we found the rate of MDS transition to AL increased with subtypes progressing(P＜0.05 ).The immune phenotype analysis of 148 patients was undertook and found that the trend to express myeloid specific antigen (CD33) increased gradually with subtypes progressing The chromosome inspection in 138 patients was undertook and found that 53 patients (38.7 ％ with abnormal karyotype,mainly in 20q- and +8;16 cases with complex abnormal karyotype (28 ％), two patients in 5q-. 180 patients were underwent bone marrow biopsy at the same time and found that 19 patients with abnormal morphology;42 patients with bone marrow fibrosis.ConclusionsCombining with multiple index to detect the MDS contributes to the classification and diagnosis more accuratcly and long-term follow-up helps to judgment the prognosis.

This study was aimed to observe the pharmacodynamics of genistein combined with tripterygium gly-cosides ( GTW ) in the rheumatoid arthritis ( RA ) treatment of ovariectomized rats . SD rats were randomly divid-ed into four groups , which were the sham-operation group , model group , Methotrexate ( MTX ) treatment group ( 2.7 mg/kg ) , and genistein ( 30 mg/kg ) combined with GTW ( 70 mg/kg ) treatment group . There were ten rats in each group . Except the sham-operation group , rats in other groups were ovariectomized and immunized by collagen type II to prepare collagen induced arthritis ( CIA ) model . The intragastric administration of drugs was given once a day for two weeks . The arthritis index , joints swelling degree , and levels of anti-Col II , IL-6 and IL-10 in serum were tested . The results showed that the arthritis index , joints swelling degree , and levels of anti-Col II , IL-6 and IL-10 in serum were all significantly increased in the model group ( P < 0 . 05 or P< 0 . 01 ) . Compared with the model group , the arthritis index and joints swelling degree of both MTX and genistein combined with GTW were significantly decreased . The level of anti-Col II in serum of the MTX treatment group was significantly decreased ( P < 0 . 05 ) . However , there were no significant changes on levels of IL-6 and IL-10 . The levels of anti-Col II and IL-6 in serum of the genistein combined with GTW were significantly decreased ( P < 0 . 05 ) . The level of IL-10 was significantly increased ( P < 0 . 01 ) . It was conclud-ed that genistein combined with GTW can significantly inhibit the pathological progress of CIA on ovariec-tomized rats . Compared with MTX , the genistein combined with GTW received a better regulating effect on the expression of IL-6 and IL-10 . The results suggested that phytoestrogen combined with GTW played a good therapeutic effect on gonadal dysfunction type of RA . It provided novel strategies and experimental evidences on postmenopausal RA treatment in the clinical practice .