ynthesis.

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.

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

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.

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 investigate the tissue localization of CD4+T cells producing IL-17,namely Th17 cells.in patients with systemic lupus erythematosus (SLE),as well as its relationship with the activity of lupus.Methods By using H&E staining.double-label immunofluorescence.immunohistochemistry and confocal microscopy.the localization of Th17 cells was carried out in peripheral blood mononuclear cells (PBMCs).affected tissue of skin and lung obtained from 4 patients with active SLE and 2 normal human controls.Flow cytometry.reverse transcription PCR.ELISA were used to detect the proportion of Th17 cells in PBMCs,the mRNA expression of interleukin-17(IL-17)A and IL-17 F,and serum level of interleukin 17,respectively,in 50 consecutive adult patients with SLE and 15 normal human controls.Results Th17 cells were detected in PBMCs of patients with active SLE.and the fuorescence intensity of IL-17 was significantly higher in patients with active SLE than in normal human controls(127.6±20.5 vs 40.6±11.1,P＜0.001).Infiltrates of Th17 cells were noted in both skin and lung tissues of patients with active SLE.but not in those of normal human controls.The proportion of Th17 cells in PBMCs was increased in patients with active SLE.and the proportion positively correlated with SLE disease activity index(SLEDAI) (r=0.725,P＜0.01).Further more.a significant increase was observed in the mRNA expression of IL-17 A and IL-17 F and serum level of IL-17 in patients with active SLE compared with normal human controls.The amount of Th17 cells was positively correlated with the development of vasculitis.and it experienced a decrease with the remission of SLE.Conclusions A proliferation of Th17 cells is noted in patients with active SLE.which seems to closely correlated with the activity of SLE and may take part in the development of vasculitis in SLE.

Objective To investigate the safe region for screw fixation in atlas lateral mass by ra- diologic measurements to conduct the operative procedure.Methods The dimensions of the pedicles were determined in 30 patients with normal atlas by CT scan and three dimensional reconstruction images of the upper cervical spine.The space available for the screw(SAS)was defined as the perpendicular distance between two lines tangential to the spinal canal and the transverse foramen,respectively.SAS was evaluated at 0?(SAS1)and 10?(SAS2)insertion angles.The location of screws in 13 cases of opera- tion were observed by CT scan images measurements of the cervical spine.Results SASI at 0?inser- tion angle was(7.81?1.28)mm and SAS2 at 10?insertion angle was(9.69?1.23)mm,with statisti- cal difference(P＜0.01).For SAS1,the distance from the midline of pedicle to the midline of posterior arch and the distance from the center of C2 inferior articular process to the midline of posterior arch showed insignificant statistical difference.However,the entry point at the posterior arch of the midline of SAS2 was located at 2.0 mm lateral to the midline of ASAI.Twenty-six pedicle screws in 13 cases were inserted correctly,without cortex breakage,or nerve and vessel injury.Conclusions There is a safe region for screw fixation in atlas lateral mass.Wider space available for the screw can be acquired when transverse angle is 10?.It is necessary to confirm the safe region by using axial CT of atlas before screw fixation.

Objective To explore the effect and safety of transcranial approach for spheno-orhital meningioma. Design Retro- spective case series. Participants Thirty-two patients being operated with transcranial approach. Twenty-four cases were meningothelial meningiomas, 3 cases were fibrous meningiomas, 1 case was psammomatous meningioma, 2 cases were atypital meningiomas, 2 case were malignant meningiomas. Methods All patients underwent frontal-temporal craniotomy, the involved sphenoid wing bone and peri- orbit were removed to prevent recurrence. The superior orbital fissure and optic canal were decompressed, the dural and periorbital de- feet were repaired by autogenous temporal fascia or artificial dura. Main Outcome Measures Preoperative and postoperative exoph- thalmus and eyeball movement, the extent of tumor resection, the ratio of recurrence. Results The extent of tumor resection: 8 cases were Simpson gradeⅡ, 20 cases Simpson gradeⅢ, 4 cases Simpson grade IV. After surgery, proptosis were improved in all patients, ophthalmoplegia was found in 6 eases. There was no operation-related death or other significant complication. Tumor recurred in 6 cas- es. Conclusions Adequate exposure of the tumor and bony decompression of the cranial nerves can result from transcranial approach, all the involved bone should be removed in order to prevent recurrence. This approach is relatively safe and the ptoptosis are improved significantly. Complete surgical resection is difficult because of the involvement of the orbital apex, superior orbital fissure and cav- ernous sinus.