OBJECTIVETo observe the effect of tetrandine on gene expression of collagen type I, collagen type III, transformation growth factor-beta1 and to investigate the inhibitory effect of tetrandine on the scar tissue hyperplasia in rabbits' ears.

METHODSAfter the scar model was formed on the rabbits' ears, the rabbits were divided into 4 groups to receive intro-lesion injection with saline, or prednisolone (Pre) or tetrandrine in low concentration (L-Tet, 1.0 mg/ml) or tetrandrine in high concentration (H-Tet, 7.5 mg/ml). The morphological changes of scar tissue were observed. The changes of fibroblasts quantity and collagen expression were observed with HE and Masson staining. Immunohistochemical study was used to observe the expression level of collagen type I and collagen type III and TGF-beta1. Collagen type I and collagen type III and TGF-beta1, and signal factor Smad 3 mRNA were detected with RT-PCR.

RESULTS(1) 24 days after injury, all the wounds healed completely with formation of red, tough and hypertrophic scar. HE and Masson staining showed significant increase of fibroblasts and collagen density with irregularly arrangement. (2) Compared with that in saline group, the scar in other groups became softer, lighter and thinner, especially in H-Tet group. (3) HE and Masson staining shows the scar in Tet and Pre groups contained less fibroblasts and lower collagen dentsity with comparatively regular arrangement than that in saline group (P < 0.01), especially in H-Tet group. (4) According to the immunohistochemical study, the expression of collage type I and III and TGF-beta was positive in all the groups, but the positive rate and the ratio of collagen density I to III decreased in the order of saline, L-Tet, H-Tet and Pre groups (P < 0.01). (5) PT-PCR detection results showed that the amplification bands brightness of collagen type I and III and TGF-beta1 and signal molecular Smad 3 mRNA in scar tissue were obviously different. Compared with that in saline group, the expression of collagen type I and III and TGF-beta1 and Smad 3 mRNA decreased in Tet and Pre groups (P < 0.01). H-Tet group showed the most obvious reduce in the expression of type I collagen and TGF-beta1 and Smad 3 mRNA. Conclusions Tetrandine can significantly suppress the expression of collagen type I and collagen type III and TGF-beta1 on hypertrophic scar of rabbit ears, and reduce signal factor Smad 3 mRNA' s expression. It may be one of the important mechanism for its inhibitory effect on scar hyperplasia.

Animals ; Benzylisoquinolines ; pharmacology ; Cicatrix, Hypertrophic ; drug therapy ; genetics ; pathology ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Ear ; Fibroblasts ; Gene Expression ; Male ; RNA, Messenger ; metabolism ; Rabbits ; Smad3 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism

AIM: To measure the macular thickness of normal young people by 3D 1000 optical coherence tomography (OCT) and study the repeatability of measuring results and the relationship between the thicknesses of macular and gender. At the same time, to compare our result with the data of other types of OCT, and to understand the consistency of the measuring results of macular thickness of different types of OCT.
METHODS: Totally 222 eyes in 111 young people were detected using 3D scan mode of Topcon 3D OCT 1000 (ver 2.4 ) . Twelve cases ( 24 eyes ) underwent repeatability check. We took transverse comparison between our measured results with other research's results.
RESULTS: There were 111 cases of young people, whose age were from 18-27 years old, all uncorrected and corrected visual acuity were≥1. 0, all intraocular pressure were <21mmHg. The average thickness of all macular region was 273. 32±17.08μm. Retinal volume of macular area was 7. 73 ± 0.37mm3 . Center thickness was 161 -264μm, and the average thickness was 200. 13±18. 81μm. Central macular thickness were 188 - 273μm, and the average thickness was 229. 00 ± 18. 20μm. The central macular thickness in men was significantly greater than that in women, and there was statistical difference. The results of repeated check of 12 cases ( 24 eyes ) in the macular area were no statistical difference except the outer ring of nasal quadrant, and the repeatability of average thickness in central macular thickness was better than in center thickness.
CONCLUSION:The repeatability of macular examination is good. The central macular thickness can be better repeated than the center thickness. The central macular thickness is 229. 00±18. 20μm in young people, according to the 3D 1000 OCT measurements. There are statistical difference of central macular thickness between different genders.

OBJECTIVETo evaluate the optimal timing of hepatectomy for intrahepatic lithiasis complicated with acute cholangitis.

METHODSOne hundred and twenty-six patients with hepatolithiasis who had a history of acute cholangitis and underwent hepatectomy were reviewed retrospectively. According to the period between the surgery and last attack of acute cholangitis, 126 patients were divided into 3 groups: > 3 months (group A, n = 73), 1 approximately 3 months (group B, n = 28), < 1 month (group C, n = 25). The operation time, blood loss, hospital stay, postoperative complications and stone residual rate were compared among the groups.

RESULTSThe intraoperative blood loss of C group was (644.0 +/- 625.7) ml, which was significantly higher than those of A and B group [(409.2 +/- 250.7) ml and (423.2 +/- 237.1) ml, respectively]. The numbers of patients who needed transfusion and the amount of blood transfusion in group C were also higher than those of group A and B. The incidence rate of complications, residual stone in group C were all markedly higher than those of group A and B. The period of hospital stay in group C was much longer than that in group A and B.

CONCLUSIONSThe optimal timing of hepatectomy for hepatolithiasis complicated with acute cholangitis is at least one month after subsidence of cholangitis.

Adult ; Aged ; Bile Ducts, Intrahepatic ; Cholangitis ; complications ; Cholelithiasis ; complications ; surgery ; Female ; Hepatectomy ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Time Factors

BACKGROUNDOsteopontin (OPN) is one kind of cytokine which can play a number of roles in promoting activation of T lymphocyte, regulating balance between Th1 and Th2, participating in cell-induced immunologic response and stimulating B lymphocyte to express multi-clone antibodies. Some researches have showed that OPN may be involved in the pathogenesis of systemic lupus erythematosus (SLE). The aim of this study was to investigate possible association of a single nucleotide polymorphism (SNP) at position 9250 in exon 7 of the OPN gene (OPN gene 9250) with SLE in Chinese patients.

METHODSTotally 158 patients (18 males and 140 females) fulfilled the revised criteria for SLE by the American College of Rheumatology in 1982 and 180 healthy volunteer controls (34 males and 146 females), all from the south of China, consented to participate in the study. OPN gene 9250 polymorphism was detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP).

RESULTSThe frequency of TT genotype of the OPN gene 9250 was significantly lower (52.5% vs 70%, P < 0.05) and the frequency of TC genotype of the OPN gene 9250 was significantly higher (43.7% vs 29.4%, P < 0.05) in SLE patients than in controls. There were significant differences in OPN gene 9250 allele and phenotype frequencies between the SLE patients and controls (P < 0.05). When the SLE patients and controls were separated into men and women, significant differences of frequencies were noted in TT genotype, TC genotype and allele of the OPN gene 9250 in women (P < 0.05) but not in men (P > 0.05).

CONCLUSIONSOPN gene 9250 polymorphism appears to be associated with susceptibility to SLE in Chinese Han ethnic population.

Adolescent ; Adult ; Aged ; China ; ethnology ; Female ; Humans ; Lupus Erythematosus, Systemic ; genetics ; Lupus Nephritis ; genetics ; Male ; Middle Aged ; Osteopontin ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length

Arrhythmogenic Right Ventricular Dysplasia ; virology ; Female ; Flavivirus ; isolation & purification ; Hepacivirus ; isolation & purification ; Humans ; Microscopy, Electron ; Middle Aged ; Myocytes, Cardiac ; virology ; Virion ; isolation & purification

OBJECTIVETo compare the difference of vasomotor functions in aortas segments from Wistar rats between 1-hour and 6-hours after exposure of water-soluble components of fine particulate matter (PM2.5).

METHODSAll 30 Wistar rats were assigned to five groups (n=6 for each group) at random: the blank control group, control group for 1-hour and 6-hours, exposure group for 1-hour and 6-hours. The rats were sacrificed 1-hour or 6-hours later and aorta ring segments were mounted on wire myographs.

RESULTS(1) There was no significant difference in vasomotor functions among three control groups (P>0.05). (2) 1-hour or 6-hours after exposure there was a decrease of contraction elicited by 60 mmol/L KCl in contrast to the control group (P<0.05), whereas no significant change between the exposure group for 1-hour and 6-hours (P>0.05). (3) On the level of 10(-5) or 10(-7) mol/L, 1-hour after exposure there was a decrease in endothelium-dependent acetylcholine (ACh) elicited relaxation precontracted by 10(-6) mol/L NE compared with the control group (P<0.01 or P<0.05), on the level from 10(-5) to 10(-7) mol/L there was a decrease compared with the exposure group for 6-hours (P<0.05), whereas no difference between the exposure group for 6-hours and the control group (P>0.05). On the level from 10(-5) to 10(-9) mol/L, 1-hour after exposure there was a decrease in endothelium-independent sodium nitroprusside (SNP) elicited relaxation precontracted by 10(-6) mol/L NE as compared with the control group (P<0.01 or P<0.05) and a decrease on the level of 10(-6) or 10(-9) mol/L compared with the exposure group for 6-hours (P<0.05), 6-hours after exposure a decrease was caused as compared with the control group on the level from 10(-5) to 10(-7) mol/L (P<0.01 or P<0.05).

CONCLUSIONSInhibition of contraction and impairment of relaxation in aortas should be caused 1-hour after exposure to water-soluble components of PM2.5 in the air, which is weaken 6-hours after exposure.

Acetylcholine ; blood ; Air Pollutants ; toxicity ; Animals ; Aorta, Thoracic ; Male ; Particulate Matter ; toxicity ; Rats ; Rats, Wistar ; Vasoconstriction ; drug effects ; Vasodilation ; drug effects

BACKGROUNDA novel anti-rheumatic drug, T-614, has been shown to have an anti-inflammatory effect and to improve abnormal immunological findings in rheumatoid arthritis (RA). To assess the safety and efficacy of T-614 versus placebo in patients with active RA we conducted a 24-week clinical study in 280 Chinese patients.

METHODSIn a multicenter, randomized, double blind, placebo controlled study, 280 patients were randomly assigned to receive placebo (n = 95) or T-614 at 50 mg (n = 93) or 25 mg (n = 92) daily. Active disease was defined by 4 of the following 5 criteria: >or= 5 tender joints, >or= 3 swollen joints, morning stiffness lasting for >or= 60 minutes, and Westergren erythrocyte sedimentation rate (ESR) >or= 28 mm/h, the assessment of pain at the rest by patient as moderate or severe. Clinical and laboratory parameters were analyzed at baseline, 2, 4, 6, 12, 18 and 24 weeks. The primary efficacy variable at week 24 was the American College of Rheumatology (ACR) response rate using the intent-to-treat population.

RESULTSThe ACR response rate was significantly higher in the T-614 treatment group compared with the placebo group within 8 weeks after the initiation of treatment. After 24 weeks, the 25 mg/d and 50 mg/d dosage groups and the placebo group showed 39.13%, 61.29% and 24.21% in ACR20 and 23.91%, 31.18% and 7.37% in ACR50, respectively. A time-response in ACR response was observed, with clear superiority for the 25 mg/d and 50 mg/d dosage groups compared to placebo (P < 0.0001), and the 50 mg/d dose compared to the 25 mg/d dose (P < 0.05) when using the ACR response analyses after 24 weeks. ESR and c-reactive protein (CRP) were significantly different in the treatment groups after 24 weeks. The incidence of adverse events (AEs) was not significantly higher with T-614 than with placebo, but upper abdominal discomfort, leucopenia, elevated serum alanine aminotransferase (sALT), skin rash and/or pruritus were more common in the 50 mg and 25 mg dosage groups.

CONCLUSIONT-614, a new slow-acting drug, is effective in treatment of rheumatoid arthritis and is well tolerated.

Adult ; Aged ; Antirheumatic Agents ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; Benzopyrans ; adverse effects ; therapeutic use ; Double-Blind Method ; Female ; Humans ; Male ; Middle Aged ; Sulfonamides ; adverse effects ; therapeutic use

OBJECTIVETo investigate the value of contrast-enhanced ultrasound in the differential diagnosis of solid focal lesions of pancreas (s-FLPs).

METHODSWe retrospectively analyzed the clinical data of 56 s-FLPs examined with contrast agent combined with low mechanical indicators contrast-enhanced ultrasound.

RESULTSThe surrounding pancreas parenchyma enhancement time, lesion enhancement time, and peak enhancement time between different groups of s-FLPs had no significant differences (P > 0.05), while the beginning to peak enhancement time showed significant difference (P < 0.05). When using the enhancement speed as a diagnostic indicator to differentiate pancreatic carcinoma from tumor-like pancreatitis, the sensitivity, specificity, and accuracy were 90.5%, 71.4%, and 85.7% for pancreatic carcinoma and 75.0%, 91.7%, and 89.3% for tumor-like pancreatitis. When using the enhancement pattern as a diagnosis indicator to differentiate pancreatic carcinoma from tumor-like pancreatitis, the sensitivity, specificity, and accuracy were 85.7%, 78.6%, and 83.9% for pancreatic carcinoma and 75.0%, 100%, and 94.6% for tumor-like pancreatitis. When different indicators were combined, enhancement pattern and enhancement speed showed the best diagnostic results; however, the Youden index was not improved.

CONCLUSIONSDifferent s-FLPs show different enhancement findings on contrast-enhanced ultrasound. The enhancement pattern and enhancement speed are the most useful diagnostic indicators.

Diagnosis, Differential ; Humans ; Pancreas ; diagnostic imaging ; Pancreatic Neoplasms ; diagnostic imaging ; Pancreatitis ; diagnostic imaging ; Retrospective Studies ; Ultrasonography

OBJECTIVETo study the regulatory rolesof SIRT1 on EZH2 expression and the further effects on EZH 2’ s repression of target gene expression.

METHODSThe stable SIRT1 RNAi and Control RNAi HeLa cells were established by infection with retroviruses expressing shSIRT1 and shLuc respectively followed by puromycin selection. EZH2 protein level was detected by Western blot in either whole cell lysate or the fractional cell extract. Reverse transcription-polymerase chain reaction was performed to detect the mRNA level of EZH2. Cycloheximide was used to treat SIRT1 RNAi and Control RNAi cells for protein stability assay. Chromatin immunoprecipitation(ChIP) assay was applied to measure enrichment of SIRT1, EZH2, and trimethylated H3K27 (H3K27me3) at SATB1 promoter in SIRT1 RNAi and Control RNAi cells.

RESULTSWestern blot results showed that EZH2 protein level increased upon SIRT1 depletion. Fractional extraction results showed unchanged cytoplasmic fraction and increased chromatin fraction of EZH2 protein in SIRT1 RNAi cells. The mRNA level of EZH2 was not affected by knockdown of SIRT1. SIRT1 recruitment was not detected at the promoter regionof EZH2 gene locus. The protein stability assay showed that the protein stability of EZH2 increases upon SIRT1 knockdown. Upon SIRT1 depletion, EZH2 and H3K27me3 recruitment at SATB1 promoter increases and the mRNA level of SATB1 decreases.

CONCLUSIONSDepletion of SIRT1 increases the protein stability of EZH2. The regulation of EZH2 protein level by SIRT1 affects the repressive effects of EZH2 on the target gene expression.

DNA-Binding Proteins ; analysis ; chemistry ; physiology ; Enhancer of Zeste Homolog 2 Protein ; Gene Expression Regulation ; HeLa Cells ; Humans ; Polycomb Repressive Complex 2 ; Repressor Proteins ; physiology ; Sirtuin 1 ; antagonists & inhibitors ; physiology ; Transcription Factors ; analysis ; chemistry ; physiology

OBJECTIVETo develop an alternative method for assessment of gene delivery systems in vivo.

METHODSMouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase (Gluc) expression cassette. After implantation of these cells into recipient mice, the expression of Gluc was detected in whole blood or plasma collected.

RESULTSAs little as 10 muL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a luminometer. And the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the mice.

CONCLUSIONSGluc may be useful as an in vivo reporter for gene therapy researches, and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.

Animals ; Arecaceae ; enzymology ; Cell Line ; Gene Transfer Techniques ; Genes, Reporter ; Humans ; Luciferases ; genetics ; Mice