OBJECTIVE:To establish HPLC method for the determination of paeoniflorin in Senqi sodium ferulate cap?sules.METHODS:The determination was performed on C 18 chromatographic column.The mobile phase consisted of acetoni?trile-water(17∶83)with a flow rate of1.0ml/min.The column temperature was30℃;the detection wavelength was230nm and the sample size was10?l.RESULTS:Good linear relationship with peak area occurred when the sample size of paeoniflorin was at a range of0.10ug～1.00ug(r=0.9999).The average recovery was98.76%(RSD=1.35%).CONCLUSION:This method is easy to handle yet with accurate results,good reproducibility and high recovery rate;and it can be used for the as?saying and quality control of Senqi sodium ferulate capsules.

BACKGROUND: The proliferative capacity of in vitro cultured chondrocytes is gradually reduced following several passages. Many studies have demonstrated that basic fibrublast growth factor (bFGF) can promote chondrocyte proliferation, and influence its phenotype and differentiation. OBJECTIVE: To observe the influence of bFGF on the proliferation and differentiation of growth plate chondrocytes and to screen the best dosage for in vitro culture of chondrocytes. DESIGN, TIME AND SETTING: For observation and comparison of cells, the experiment was performed at Tissue Engineering Laboratory, Department of Biochemistry, Jinan University Medical School between October 2004 and February 2005. MATERIALS: Twelve New Zealand rabbits were selected for chondrocyte isolation; bFGF was purchased from PeproTech, UK. METHODS: Rabbit growth plate chondrocytes were isolated and cultured in low serum. Different doses of bFGF (0, 1, 2.5, 5, 10, 25, 50 and 100 μg/L) were added. MAIN OUTCOME MEASURES: Cell proliferative multiple was determined using modified MTT method; chondrocyte collagen content was determined using hydroxyproline method; alkaline phosphatase activities were detected by enzyme kinetics. RESULTS: bFGF (5-100 μg/L) could promote chondrocyte proliferation, especially at dose of 25 μg/L. bFGF at dose of larger than 25 μg/L would inhibit collagen production; bFGF at dose of larger than 1 μg/L would inhibit alkaline phosphatase activities. CONCLUSION: bFGF can stimulate growth plate chondrocyte proliferation, and inhibit growth plate chondrocyte differentiation at dose25 μg/L.

82 cases were treated with acu—moxibustion and tra-ditional materia medica,while 42 cases were treatedwith routine western drugs.Results revealed that theamelioration of clinical symptoms,signs and total ef-fectiverate in the traditional modality group was betterthan the western drug group.

Objective To investigate the value of transoral carotid ultrasonography (TOCU) in detecting the change of distal extracranial internal carotid artery(ICA) with stenosis and occlusion in its initial segmen.Methods One hundred and ten patients with stenosis at least 50％ or occlusion in the initial segment of ICA diagnosed by high-frequency linear probe were enrolled as case group,while thirty two with healthy carotid arteries as control group.Both of the case and control groups were undergone high-frequency linear probe to examine CCA and the initial segment of ICA,and TOCU to observe their lumen transparency,color Doppler flow display and change of Doppler flow spectrum,even measuring the internal diameter,peak systolic velocity (PSV),end-diastolic velocity(EDV) and PSVICA/PSVCCA ratio,respectively.Digital subtraction angiography (DSA)were performed within two weeks,then compared with ultrasonography.Results 92.7％ patients were performed TOCU examination in case group and 93.8％ in control group.The internal diameter and PSV of the distal extracranial ICA became gradually smaller from control group to near occlusion group,and there were statistical significance between groups ( P ＜0.05),however,near occlusion group had no statistical significance compared with occlusion group( P ＞0.05).There were 80％ arteries having thrombus echo in occlusion group,and the others in this group having no thrombus had gloomy colour flow,even got low-velocity artery spectrum.Doppler flow spectrum of distal extracranial ICA showed characteristics of slow wave or single peak in near occlusion group,and spike shape or slow wave in 76.7 ％ arteries of 70％ ～near-occlusion stenosis group,but spike shape in 30.8％ arteries of 50％～69％ stenosis group,and the others were normal.Conclusions TOCU is useful for detecting the changing regularity of distal extracranial ICA with stenosis or occlusion in its initial segment and has the value of clinical application in association with high-frequency linear probe.

ObjectiveTo assess the feasibility of evaluation of renal allograft acute rejection in rat withcontrast-enhancedultrasound( CEUS )andtargetedmicrobubbles.MethodsPhospholipid microbubbles targeted to intercellular adhesion molecule -1 (ICAM-1)(MBI) and control microbubbles (MB) were created by conjugating monoclonal antibody against ICAM-1 or isotype control antibody to the lipid capsule via “avidin-biotin” bridging.Ten SD rats with acute renal allograft rejection were injected intravenous of MBI and MB in random order with a 30-min interval.After 3 min of intravenous injection of microbubbles,targeted CEUS imaging was performed in all rats.And then the video intensity (VI) was determined.ResultsIn MBI group,a significant ultrasonic enhancement was observed,but it was not very obvious in MB group.Increment in VI value of transplant kidney in MBI group was great and it amounted to (27.0 ± 7.4)U,however,increment in VI value of in MB group was minor and it was merely (10.2 ± 2.4) U,Difference was evident in transplant kidney between of the two groups (P ＜ 0.05).Conclusions Molecular imaging of ICAM-1 with targeted CEUS can evaluate renal allograft acute rejection effectively.

AIM To investigate the expression of pep-tide transporter (PEPT)2 mRNA in the lungs of ratswith lipopolysaccharide(LPS)-induced acute lung inju-ry. METHODS Health male Sprague-Dawley rats were divided randomly into 5 groups: normal control,cheally) given groups. Lung histopathological changes were observed under light microscope. Lung wet-to-dry weight (W/D) ratios, total protein concentration in bronchoalveolar lavage fluid (BALF) and lung my-eloperoxidase(MPO) activity were measured. The ex-pression levels of PEPT2 mRNA were evaluated by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS Typical pathological inflammation changes in the lungs, including alveolar congestion, hemorrhage, edema, infiltration of neutro-phils in the air space or vessel wall, thickness of the alveolar wall, and hyaline membrane formation, were observed in LPS groups. The lung W/D ratios in LPS 2, 4 and 8 h groups (4.72±0.18, 5.06±0. 17 and 5.12±0.16) were significantly higher than those innormal control and saline groups (4.12±0. 15 and 4.14±0.11). Compared with normal control and sa-g-1 wet tissue], MPO activity increased obviously in LPS2,4 and 8 h groups[(1.96±0.15), (2.23±protein concentration in BALF in LPS 2, 4 and 8 hgroups [ (36.6±2.9), ( 86.9±3.5 ) and (92.2±There was no statistical difference in PEPT2 mRNA levels in the lungs of rats among all groups. CON-CLUSION PEPT2 mRNA level does not change sig-nificantly in the acute injured lung tissue induced by LPS.

Objective To investigate the incidence of subclinical hypothyroidism (SCH) during the third trimester of pregnancy and its effects on pregnancy outcomes and neonatal hypothyroidism.Methods A total of 10 695 women in the third trimester of pregnancy (28-42 weeks of gestation) who labored from January 1,to December 31,2012 in Changzhou Maternal and Child Health Care Hospital Affiliated to Nanjing Medical University were enrolled.The levels of thyroid stimulating hormone (TSH),free thyroxine (FT4) and thyroid peroxidase antibody (TPOAb) of the mothers were quantified by electrochemical immunoassay.The time-resolved fluorescence immunoassay was used to detect neonatal thyroid hormone levels.Using t test or x2test,the incidence of adverse pregnancy outcomes was compared between SCH group and euthyroid group and between SCH women with positive (n=40) or negative TPOAb (n=176).Results The incidence of SCH was 2.02％ (216/10 695) and the positive rate of TPOAb in SCH women was 18.5％ (40/216).No neonatal thyroid dysfunction was found.According to the age matched,222 cases were randomly selected as controls from 7 757 euthyroid women.Compared with the controls,SCH women had a higher incidence of premature rupture of membranes [28.7％ (62/216) vs 14.9％ (33/222),x2=12.34],anemia [11.6％ (25/216) vs 4.1％ (9/222),x2=8.65],pregnancy-induced hypertension [9.7％ (21/216) vs 4.5％ (10/222),x2=4.53],premature labor [8.8％ (19/216) vs 3.6％ (8/222),x2=5.10] and intrahepatic cholestasis of pregnancy [8.3％(18/216) vs 2.3％ (5/222),x2=8.14] (all P＜0.05).The overall incidence of adverse pregnancy outcomes was also higher in SCH group than in the controls [69.4％ (150/216) vs 49.5％ (110/222),x2=17.96,P＜0.01].The incidence of fetal growth restriction and still birth in SCH mothers with positive TPOAb was higher than in those with negative TPOAb [7.5％ (3/40) vs 0.0％ (0/176),x2=13.32,P＜0.01; 2.5％ (1/40) vs 0.0％ (0/176),x2=4.40,P＜0.05],but there was no significant difference in the overall incidence of adverse pregnancy outcomes compared with TPOAb-negative mothers [65.0％ (26/40) vs 70.5％ (124/176),x2=0.46,P=0.50].Conclusions SCH diagnosed in the third trimester may lead to adverse pregnancy outcomes.Early screening for thyroid dysfunction is necessary.

Objective To study the effects of psychological stress on breast development and abnormal levels of related hormones or their receptors in pregnant rats .Methods Pregnant Wistar rats were randomly divided into control group and ex-perimental group.The experimental group rats were given unpredictable stress stimuli for 15 days, including noise, restraint, re-versal of day and night , swimming in cold water and tail pinch-induced pain.Gross changes of the breast were observed , the di-ameter and height of breast nipple was measured, and breast coefficient was calculated.DNA and RNA levels in the breast tis-sues were determined by optical density measurement.The estrogen (E2), progesterone (P), growth hormone (GH) and prolac-tin (PRL) levels in plasma and breast tissues were determined by radioimmunoassay.Bmax and Kd of estradiol and progesterone receptors were measured by radolig and bindng assay .Morphological changes of the breast tissues were examined by light micros-copy using HE staining.Results Compared with the control group, the nipple diameter and height, breast weight and breast co-efficient were significantly decreased in the experimental group (P<0.05, P<0.01), the levels of DNA, RNA and RNA/DNA ratio in the breast tissue were significantly decreased (P<0.05, P<0.01), the levels of plasma E2, P and GH were significant-ly decreased (P<0.05, P<0.01), the levels of E2 and GH in the breast tissue homogenate were decreased also (P<0.05, P<0.01), the Bmaxs of E2 receptor and P receptor were decreased, but their Kds increased significantly (P<0.05, P<0.01), and the number of lobules and diameter of acini were significantly decreased (P<0.05).Conclusions Psychological stress can lead to breast dysplasia and abnormal levels of estrogen and progesterone and their receptors in pregnant rats .

Objective To investigate the effects of ultrasound radiation force(USRF) on adhering of targeted microbubbles in vivo.Methods A rat model of cremaster muscle Inflammation was developed,and microbubble(MB) or targeted microbubble of intercellular adhesion molecule-1 (MBICAM) was injected through the caudal vein.Immediately after intravenous MB or MBICAM injection,the cremaster was insonated for 5 minutes and the control group was in sham ultrasound exposure.All mice were divided into four groups randomly:① MB + false USRF; ② MB + USRF; ③ MBICAM + false USRF; ④ MBICAM + USRF.Fluorescence microscope was performed in twenty mice to evaluate microbubbles adhesion and video recordings were made with a high-resolution camera.Results The adhesion number of MBICAM was significantly greater in USRF group than that of MBICAM in false USRF group [(43.4 ± 2.1)/view vs (14.8± 1.8)/view,P =0.000].The adhesion number of MB in USRF group was greater than in false USRF group [(6.2 ± 1.3)/view vs (4.6 ± 0.9)/view].But there is no statistical significance (P =0.129).The adhesion number of MBICAM in both USRF group and false USRF group was significantly greater than the adhesion number of MB.Conclusions The ultrasound radiation force can significantly improve the adhesion of targeted microbubbles in vivo.

Objective To construct a recombinant plasmid for expressing the HPV18 E2 gene and develop some materials for HPV18 related disease.Methods The E2 gene of HPV18 was amplified by PCR from PBR322-HPV18 and cloned into PIRES2-EGFP.The sequence of cloned HPV18E2 was confirmed by restriction analysis and DNA sequencing,then transfected into HeLa cells.E2 gene was detected by RT-PCR.Results HPV18 E2 gene fragment was amplified by PCR and ligated to PIRES2-EGFP,then transfected into HeLa cells successfully.The expression of green fluorescence cells was observed by fluorescence microscopy.Specific band could be seen in transfected cells by RT-PCR.Conclusion Recombinant plasmid PIRES2-EGFP is effectively expressed after being transfected into HeLa cells in vitro.