Objective To study the possible relationships between expression of matrix metalloproteinase(MMP)2,9 and the pathogenesis of preeclampsia in which trophoblast invasion is impaired. Methods MMP-2,9 expression were detected by immunohistochemistry streptavidin-biotin complex (SABC)method in 20 normal term placentae and 20 preeclampsia placentae,respectively.In addition, mRNAs for MMP-2,9 were analyzed by real time PCR in both groups.Results The intensities of both MMP-2 and MMP-9 immunostaining in preeclampsia placentae were significantly declined compared to those of normal term placentae(P

Objective To test the hypothesis that homocysteine can decrease MMP-2 and MMP-9 expression in cultured trophoblasts of early pregnancy and that homocysteine can prevent trophoblasts invasion in the early stage of preeclampsia.Methods Cytotrophoblasts from early pregnancy were isolated and cultured.Trophoblasts were treated with or without Hcy(1 mmol/L)for 48 hour,and real time RT-PCR and gelatin zymography were used to quantify the mRNA and protease activity of MMP-2,-9.Results Treatment with Hcy(1 mmol/L)induced a decrease in MMP-2 mRNA by 21% and MMP-9 mRNA by 11%.At protein level MMP-2 expression decreased 14% and MMP-9 expression decreased 52% compared with control.Conclusions Homocysteine can decrease MMP-2,-9 expression in trophoblasts of early pregnancy and influence its invasion process.

A quantitative analysis method of multi-components with a single marker (QAMS) for simultaneous determination of six marker compounds (one from phenolic acids and five from phthalides) in Chuanxiong Rhizoma was established by applying HPLC and using butylidenephthalide as the internal reference substance. And also the feasibility and accuracy of the established method for quality evaluation and application of Chuanxiong Rhizoma were investigated and validated. The analysis was performed with the mobile phase consisting of acetonitrile - 0.2% aqueous formic acid. The flow rate was 1.0 mL . min-1 and the column temperature was maintained at 30 °C. The detection wavelengths were set at 252 nm (for ferulic acid, Z-ligustilide, and butylidenephthalide) and 266 nm (for senkyunolide I, senkyunolide A, and coniferyl ferulate), separately, and 20 µL was injected for analysis with gradient elution. The results showed that there were no significant differences observed between the HPLC-QAMS method and the external standard method (RSD <5%). The relative correction factors were credible (RSD < 5%) in changed chromatographic conditions. The established HPLC-QAMS method can be accurately used for simultaneously evaluating and controlling the quality of Chuanxiong Rhizoma with multi-components.
4-Butyrolactone ; analogs & derivatives ; Acetonitriles ; Benzofurans ; Chromatography, High Pressure Liquid ; Coumaric Acids ; Drugs, Chinese Herbal ; analysis ; standards ; Hydroxybenzoates ; Quality Control ; Rhizome ; chemistry

Objective To investigate the dosimetric influence of dwell weight standard deviation (DWSD) and applicator displacement in cervical cancer patients treated with three-dimensional brachytherapy.Methods A total of 20 cervical cancer patients who had completed radical treatment were selected in this study.The Fletcher applicator (Nucletron#189.730) was used for these patients.A new plan,based on the former CT images and structures,was designed for each patient.In former and new plans,dwell weight was recorded,and DWSD was calculated.Two groups,low-DWSD (LDWSD,0.141-0.299) and high-DWSD (HDWSD,0.211-0.337),were set according to the DWSD size for the two plans.Dosimetric effects from ± 1 mm displacement of tandem applicator or ovoid applicator were simulated with Oncentra (R) Brachy V4.3 treatment planning system.D100,D90,and V150 for clinical target volume (CTV)and D0.1cc,D1cc,and D2cc for the bladder,rectum,and sigmoid were evaluated.Dosimetric comparisons were made between the LDWSD group and HDWSD group to study the dosimetric effects of DWSD and applicator displacement in cervical cancer patients.Results The dosimetric effects from applicator displacement increased with increasing DWSD.If there was a 1 mm displacement of tandem applicator or ovoid applicator,D100,D90,and V150 of CTV were 3.0％,23.8％,and 4.8％ higher or 0.5％,1.2％,and 5.2％ higher in the HDWSD group than in the LDWSD group;D0.1cc,D1cc,and D2cc of the bladder and rectum were significantly higher in the HDWSD group than in the LDWSD group,particularly for the sigmoid (up 44.0％,22.8％,and 16.8％) and (up 10.3％,14.4％,and 12.4％).Conclusions DWSD should be considered in plan evaluation for cervical cancer patients treated with three-dimensional brachytherapy.The dosimetric influence from applicator displacement can be decreased by reducing DWSD properly.

The aim of the present study was to investigate the effect of lipoxin A4 (LXA4)pretreatment on cognitive function of aged rats after global cerebral ischemia reperfusion,and to explore its possible mechanism.Thirty-six aged male Sprague-Dawley rats were randomly divided into three groups (n=12 each):sham-operation group (S group),global cerebral ischemia reperfusion group (I/R group) and LXA4-pretreatment group (L group).The rat model of global cerebral ischemia reperfusion was established by occlusion of the bilateral common carotid artery with hypotension.The cognitive function of rats was determined by a step-down type passive avoidance test and Morris Water Maze test on the third day after reperfusion.Rats were sacrificed after Water Maze test and the pathological changes ofhippocampal CA1 region were observed and the related inflammatory mediators were determined.As compared with S group,the escape latency in I/R group was prolonged from the first day to the fifth day,while that in L group was prolonged from the first day to the third day.The retention time in I/R group and L group in the first quadrant was shortened.The reaction time,frequency of reaction mistake and frequency of escape mistake in I/R group increased,and the latent period shortened.The frequency of escape mistake in L group increased,and the damage in the hippocampal CA1 region of I/R group and L group was obvious.The levels of S-100β,TNF-α,IL-lβ,IL-10 and NF-κB in I/R group and L group increased.As compared with I/R group,the escape latency in L group was shortened from the first day to the fifth day,and the retention time in the first quadrant prolonged.The reaction time,frequency of reaction mistake and frequency of escape mistake in L group decreased,and the latent period prolonged.The damage in the hippocampal CA1 region of L group was alleviated as well.The levels of S-100β,TNF-α,IL-1β and NF-κB in L group decreased,and those of IL-10 increased.It can be concluded that LXA4 pretreatment can improve the cognitive function in aged rats after global cerebral ischemia reperfusion probably by inhibiting the inflammatory reaction.

Objective To observe the effect of T-2 toxin on growth and development of rat epiphyseal plate of left knee and metaphyseal bone of femur and tibia in normal and low nutritional status, to find out possible pathogenic factors of Kashin-Beck disease and provide experimental basis for early intervention. Methods Ninety 3-week-old Wistar rats, weighing 60 - 70 g, were randomly divided into three groups: control group(general feed), T-2 toxin + general feed group, T-2 toxin + low nutrition feed group, thirty rats in each group with equally sex ratio. T-2 toxin (1.0 mg/kg) was administered orally 5 times a week via a gavage needle for 4 weeks. The change of hair, activity and body weight was observed. After 1, 2, 4 weeks, the epiphyseal plate of left knee and metaphyseal bone of femur and tibia (including distal femur and proximal tibia) were collected. Specimens were processed with HE and Masson staining. The morphology of chondrocytes and matrix collagen content in epiphyseal plate was observed. Trabecular bone volume fraction in tibial metaphyseal bone was analyzed by Image-Pro Plus 6.0 software. Results In the control group, rats were in good movement and hair with light, but in T-2 toxin + general feed group and T-2 toxin + low nutrition feed group, rats were found with reduced activities and hair with dark color. Body weights(g) of the control group, the T-2 toxin + general feed group and the T-2 toxin + low nutrition feed group were 81.0 ± 6.2, 79.0 ±5.1, 77.0 ± 7.5, respectively, by the end of first week; 101.8 ± 6.7, 97.0 ± 6.8, 93.0 ± 5.3, respectively, by the end of second week; 151.1 ± 15.7, 126.5 ± 11.9, 106.5 ± 11.5, respectively, by the end of fourth week. There was significant difference in groups by second week and the fourth week (F = 9.72, 41.65, all P ＜ 0.05 ). There was significant difference among multi-groups by the fourth week(all P ＜ 0.01 ). Under light microscope, at the second weeks, coagulative necrosis of chondrocytes was found in hypertrophic zone in the two groups with T-2 toxin; at the fourth weeks, cell necrosis increased. Masson staining showed collagen staining in the two groups with T-2 toxin significantly turned to clear pale coloration, indicating that the collagen matrix was significantly reduced. Image analysis showed there was significant difference in groups at the second and fourth week(F= 9.72, 41.65, all P＜ 0.05)in tibial metaphyseal trabecular bone volume fraction. There was significant difference between T-2 toxin + low nutrition feed group[(0.55 ± 0.12)％, (0.21 ± 0.0)％] and control group[(0.67 ± 0.09)％, (0.51 ± 0.14)％] by the second and fourth week(all P ＜ 0.01 ). Conclusions Under normal nutritional status, T-2 toxin can induce hypertrophic epiphyseal cartilage necrosis, collagen content decreased in epiphyseal plate, metaphyseal trabecular bone formation disorders; in the low nutritional status, T-2 toxin can lead to rat epiphyseal necrosis and significant metaphyseal bone disorder, but whether the performance is related to Kaschin-Beck disease needs to be studied further.

Background The diagnosis and treatment of fungal keratitis are knotty.There is no quantitative method to identify the disease and judge the therapeutic effect of the antifungal agent.Studies have determined that serum (1,3-) β-D-glucan level can sensitively and specifically reflect the state of systemic mycotic-causing diseases.However,whether (1,3-) β-D-glucan level in tear can monitor and diagnose mycotic keratitis is unclear.Objective Purpose of this study was to investigate the change of tear (1,3-) β-D-glucan level following the administration of antifungal drug in fungal keratitis patients,and evaluate the diagnosis and monitor value of (1,3-) β-D-glucan in tears for fungal keratitis.Methods Sixty patients who were diagnosed as fungal keratitis by fungal culture were analyzed in Affiliated Hospital of Qingdao University Medical College from July 2010 to May 2012.The patients received the topical administration of antifungal drug for 28 days.Thirty healthy volunteers without eye disease served as normal controls.The tear of 50 μl was collected from each subject for the detection of (1,3)-β-D-glucan before the therapy,7,14,28 days after therapy and 7 days,14 days after the drugs were stopped,respectively.The dynamic changes of (1,3-) β-D-glucan levels in tears were evaluated and compared with the manifestation of the lesions under the laser scanning confocal microscope.The patients without hyphal by the laser scanning confocal microscopy and tear (1,3-)β-D-glucan level less than 20 ng/L were subsequently treated for another 7 days,and the following-up duration was 2 months.The informed consent was obtained before any medical examination was performed from each subject.Results (1,3-)β-D-glucan level in tears (Log value) was (6.37 ±0.48)ng/L in the patient group,and was significantly higher than (2.00±0.31) ng/L in the normal control group (t =2.89,P＜0.01).The lesion was smaller with the gradually clear border,and the number of mycelia was decreased under the laser scanning confocal microscope 7 days after treatment.(1,3-) β-D-glucan level in tears was gradually declined in a time-dependent manner after treatment.The (1,3)-β-D-glucan level in tears (Log) was (5.19 ± 0.42),(4.16 ± 0.33),(2.99 ±0.42),(2.91 ±0.39),(2.80±0.40) ng/L 7,14,28 days after treatment,and 7 days,14 days after the drugs were stopped,respectively,with a statistically significant difference in comparison with (6.37±0.48)ng/L before treatment (P＜0.01).(1,3)-β-D-gluean level in tears remained a lower level till the end of follow-up,and no recurrence of lesion was found in the patient group.Conclusions Detecting (1,3)-β-D-glucan level in tears is of good diagnosis and monitor value in the evaluation of fungal keratitis.

Objective To judge method performance of routine biochemistry parameters on Roche Modular PPI testing system by using westgard's method evaluation decision chart.Methods We assessed imprecision(CV)from internal quality control and inaccuracy(bias)from external quality control evaluation.Combined estimates of imprecision and inaccuracy by plotting imprecision as the x-coordinate and inaccuracy as the y-coordinate to locate an expected operating point of every item on the chart.By comparing this operating point with allowable total errors(TEa),we can decide whether the performance is acceptable or not.Results In the 27 different parameters tested,imprecision and bias of calcium were 0.08 mmol/L and 0.06 mmol/L respectively,its performance was marginal.The imprecision of creatinine,urea,glucose, sodium,chloride and phosphorus were 3.20%,2.13%,1.52%,0.89 mmol/L,1.10% and 1.55%,the bias were 4.79%,0.96%,4.63%,0.80 mmol/L,1.74% and 4.13% respectively,their performance was good.M1 other 20 items were of excellence performance.Conclusions Routine biochemistry parameters on Roche Modular PPI testing system possessed good precision and accuracy,and their performance were acceptable.To judge method performance of biochemistry testing system by using westgard' s method evaluation decision chart was easy to do and suited for clinical laboratory.

OBJECTIVETo evaluate the performance of BNII auto-analyzer system in detecting C3 and C4.

METHODSCLSI protocols (EP15-A, EP6-A, EP9-A2) and other relevant literatures were use to or evaluate the precision, accuracy, linearity of C3 and C4 detection by the auto-analyzer system, and the results were compared with the recognized standards.

RESULTSThe relative bias of C3 and C4 was less than one third of the CLIA'88 standard and the precision met the clinical requirement. The results tested by DADE BNII system were not compatible with those by Roche Modular System. C3 showed good linearity in the tests (R2>0.975, P<0.05) with a linearity range of 0.18-5.1 g/L. The linearity of C4 was not available because of lack of high-level samples.

CONCLUSIONThe performances of DADE BNII System basically meet the recognized standards in clinical detection of C3 and C4, but the method comparison needs further validation.

Autoanalysis ; methods ; Blood Chemical Analysis ; instrumentation ; methods ; Complement C3 ; analysis ; Complement C4 ; analysis ; Humans ; Nephelometry and Turbidimetry ; instrumentation ; methods ; Proteins ; analysis ; Sensitivity and Specificity

OBJECTIVETo investigate the effect of growth hormone (GH) on penile erection after reconstruction of cavernous nerves using sural nerve as an interposition nerve graft in rats.

METHODSTwenty-four male Sprague-Dawley rats (3-4 ms of age and 300-400 g in weight) were randomly divided into 2 groups: nerve graft group and GH group, each electrostimulated to determine the erectile potency 2 and 4 months after nerve graft (followed by hypodermic GH injection). The nNOS-positive nerve fibers in the corpora cavemosa were examined by streptavidin-peroxidase immunohistochemistry technique (SP method). Image analysis was used to calculate the area stained in pixel.

RESULTSElectrostimulation at 2 months produced 31.25% of erections in the GH group but none in the grafted rats. There was a significant difference in the erection rate produced by electrostimulation between the two groups at 2 months (P < 0.05). The pixel of the expression of nNOS-positive nerve fibers in the GH group (38971 +/- 7692) was also greater than that of the graft group (16538 +/- 3179, P < 0.05). At 4 months, 43.75% of the graft group and 75% of the GH group produced erections upon electrostimulation, with no significant difference between the two groups (P > 0.05). The pixels of the expression of nNOS-positive nerve fibers were 79276 +/- 12,021 and 91348 +/- 18965, respectively (P > 0.05).

CONCLUSIONGH can accelerate the regeneration of cavernous nerves after bilateral nerve grafting, and GH administration may present a new physiological approach to the treatment of erectile dysfunction after radical pelvic surgery.

Animals ; Growth Hormone ; pharmacology ; Male ; Nerve Regeneration ; drug effects ; Nitric Oxide Synthase ; analysis ; Penile Erection ; drug effects ; Penis ; enzymology ; innervation ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sural Nerve ; transplantation