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In the present paper, a new improved method for measuring NO2 concentration of atmosphere by using differential optical absorption spectroscopy of solar spectrum was reported. The sample solar spectra were collected in Chengdu city and Xiling Mountain respectively by a CCD optical multi-channel analyzer. With the upper atmosphere solar spectrum as the reference spectrum, by fitting the sample spectrum and the reference spectrum using least-square method in five spectrum ranges, the various NO2 concentrations (vertical column density) of 3.1 x 10(16) to 5.8 x 10(16) (molecule x cm(-2)) in Chengdu city and 0.7 x 10(16) to 1.0 x 10(16) (molecule x cm(-2)) in Xiling Mountain were obtained respectively. The NO2 concentrations of atmosphere showed an increasing tendency with the rising temperature.

Objective: To explore the mutations of MEF2A gene in Chinese patients with coronary artery disease(CAD)., Methods: With polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA direct sequencing, the mutation analysis of exon 11 of MEF2A gene was performed to 156 patients with CAD and 93 normal controls., Results: By DNA sequence analyzing the samples of abnormal mobility shift of SSCP, the MEF2A gene mutations were found in three patients with CAD. One of mutations was 147130(C>A)(P431Q), and the second one was 21 bases deletion(147108-147128) which was leading to the absence of 7 amino acids (424QQQQQQQ430), and the third was 147191(G>T). Three mutations were all found in one patient, but meanwhile 21 bases deletion was found in the other two patients., Conclusion: Mutations in exon 11 of MEF2A gene exist in the patients with CAD, and the mutations may be pathological.

A new method and technology for measuring time-dependent spectra excited by pulse laser was reported in the present paper, in which a multi-fiber delayer combined with a two dimensional CCD array detector was employed for collecting and processing the spectra. The multi-fiber delayer consists of five fibers with a length difference of 1-2 m (relative delay time 5-10 ns), and the optical multi channel analyzer is PARC OMA-4. The time-dependent spectra, i. e. the spectra of R6G dye laser, the SRS spectra of acetone, and the plasma emission spectra of metal Mo excited by a double frequency YAG pulse laser with a time resolution of 5 ns were obtained.

Objective: To investigate the phenomenon and influencing factors of aspirin resistance (AR) in patients taking small dose of aspirin., Methods: Three hundred and twenty-eight patients with stable cardiac and cerebral vascular diseases, diabetes mellitus, et al taking aspirin 100 mg/d for > or =14 days, and then their blood samples were collected for determination of optical platelet aggregation index using arachidonic acid (AA) and adenosine diphosphate (ADP). AR was defined as a state in which aggregation of > or =20% with AA and that > or =70% with ADP was found. Aspirin semi-resistance (ASR) was defined as meeting one of the above criteria. If both above criteria were not met, the condition was defined as aspirin sensitive. The difference in clinical characteristics among the groups and independent risk factors associated with AR and ASR were analyzed., Results: Of 328 patients, 4.9% were AR, 27.4% were ASR. Among AR+ASR group, female, elderly, diabetic and hypertensive patients were predominant, but less common in smokers. Logistic regression analysis showed that diabetes mellitus was an independent risk factor of AR and ASR [odds ratio (OR)=0.953, 95% confidence interval (CI) 0.323-0.876, P=0.013], and hypertension was independently associated with AR and ASR (OR=0.610, 95%CI 0.376-0.991, P=0.046). The risk of AR and ASR was increased in non-smokers (OR=2.231, 95%CI 1.182-4.210, P=0.013)., Conclusion: The incidence rate of AR in patients taking small dose of aspirin was 4.9%. Diabetes mellitus and hypertension are relative risk factors of AR and ASR. The risk of AR and ASR in the no-smoking patients is increased.

Mushroom polysaccharides have been shown to regulate glucose metabolism. Using male Wistar rats injected with saline (normal rats), streptozotocin (STZ-NT rats), or streptozotocin plus nicotinamide (STZ+NT rats), we investigated the hypoglycemic activity of orally ingested fruiting bodies (FB), submerged culture biomass (CM), or the acidic polysaccharide glucuronoxylomannan (GXM) of Tremella mesenterica, an edible jelly mushroom. Our results demonstrated that FB ingestion significantly attenuated the elevated blood glucose levels in an oral glucose tolerance test (OGTT) in STZ-NT rats. However, in STZ+NT rats, FB, CM, and GXM ingestion significantly attenuated the increases in food and water intake, 2-h postprandial blood glucose concentrations, and blood glucose levels in OGTT. Moreover, FB and GXM ingestion significantly decreased serum concentration of fructosamine in STZ+NT rats. Our results indicated that T. mesenterica might be developed as a potential oral hypoglycemic agent or functional food for diabetic patients and for persons with high risk for diabetes mellitus.

Pollution by polycyclic aromatic hydrocarbons(PAHs) is widespread due to unsuitable disposal of industrial waste. They are mostly defined as priority pollutants by environmental protection authorities worldwide. Phenanthrene, a typical PAH, was selected as the target in this paper. The PAH-degrading mixed culture, named ZM, was collected from a petroleum contaminated river bed. This culture was injected into phenanthrene solutions at different concentrations to quantify the biodegradation process. Results show near-complete removal of phenanthrene in three days of biodegradation if the initial phenanthrene concentration is low. When the initial concentration is high, the removal rate is increased but 20%-40% of the phenanthrene remains at the end of the experiment. The biomass shows a peak on the third day due to the combined effects of microbial growth and decay. Another peak is evident for cases with a high initial concentration, possibly due to production of an intermediate metabolite. The pH generally decreased during biodegradation because of the production of organic acid. Two phenomenological models were designed to simulate the phenanthrene biodegradation and biomass growth. A relatively simple model that does not consider the intermediate metabolite and its inhibition of phenanthrene biodegradation cannot fit the observed data. A modified Monod model that considered an intermediate metabolite (organic acid) and its inhibiting reversal effect reasonably depicts the experimental results.

Objective: To study the expressions of CD55 and CD59 in patients with hyperlipidemia and the effects of atorvastatin on it, and to identify the possible influential factors., Methods: We selected 67 patients with hyperlipidemia, and 24 healthy people matched in terms of age, sex and body weight as control. The expressions of CD55 and CD59 on white blood cells were detected by flow cytometry, and their relationships to blood lipids, complement activation indexes (C(5a), sC(5b-9)), inflammatory factors (high sensitivity C-reactive protein (hsCRP), TNF-alpha, IL-6 were analyzed. 24 patients with hyperlipidemia were treated with atorvastatin for 8-12 weeks and the expressions of CD55 and CD59 were measured before and after atorvastatin therapy., Results: The mean fluorescence intensity (MFI) of CD55 lymphocytes and monocytes were decreased in patients with hyperlipidemia compared with control (2.07 +/- 0.28 vs 2.29 +/- 0.44 and 3.45 +/- 1.02 vs 4.33 +/- 2.32, P < 0.01 and P < 0.05, respectively). CD55 positive lymphocyte MFI was negatively correlated with waist circumference, waist-hip ratio, hsCRP and C(5a). C(5a) was negatively correlated with the MFIs of CD55 positive lymphocytes, monocytes, granulocytes, and positively with TG and diastolic blood pressure. After atorvastatin therapy, the MFIs of CD59 positive lymphocytes, monocytes and granulocytes increased (4.34 +/- 1.16 vs 3.69 +/- 0.76, 4.52 +/- 1.36 vs 3.91 +/- 0.89, 5.67 +/- 1.72 vs 4.56 +/- 1.03, P < 0.05, < 0.05 and < 0.01 respectively), which were not correlated with changes of blood lipids., Conclusions: The expression of CD55 is down-regulated in hyperlipidemia, which might be influenced by obesity, abdominal distribution of adipose tissue and inflammatory status of hyperlipidemia, but not by blood lipids. The expression of CD55 is related with complement activation; The expression of CD59 is up-regulated after atorvastatin treatment independently of blood lipids.

We studied the effects of Chinese traditional medicine rhynchophylline (Rhy) on human ether-a-go-go related gene (HERG) channel and characterized the electrophysiological properties of Rhy's pharmacological effect on HERG channel using Xenopus oocytes. Xenopus oocytes were injected with either 23 nl (5.75 ng) HERG cRNA or 23 nl distilled water. Xenopus oocytes were randomly assigned to receive one of the following different concentrations of Rhy: (1) control, (2)10 mumol/L Rhy, (3)100 mumol/L Rhy, (4) 500 mumol/L Rhy, (5) 1 000 mumol/L Rhy, (6) 10 000 mumol/L Rhy. Cell currents were recorded in oocytes. The peak tail currents of HERG channel were inhibited by Rhy. The inhibition was in a dose-dependent manner [IC(50)=(773.4 +/- 42.5) mumol/L]. Experiment with 100 mumol/L Rhy indicated that the degree of HERG blockade showed some voltage dependence (within -40 mV to -20 mV ). Kinetic analyses revealed that Rhy decreased the rate of channel activation. The findings indicate that Rhy inhibits HERG encoded potassium channels. It may underline the molecular mechanism of myocardial electrophysiological characteristics associated with this drug.

A flow-injection-CCD array detection spectrophotometry for the determination of oligomeric proanthocyanidin (OPC) in health foods was developed in the present paper, based on the reaction that OPC can form red anthocyanidin in hydrochloric acid medium with ferric ion as a catalytic agent. The absorption spectrum of OPC was scanned and the absorbance at 545 nm was determined by flow injection with CCD-array detector. The influencing factors of the chromogenic reaction and parameters of the flow injection analysis were optimized. There is a good linear relationship between the absorbances and the concentrations of OPC in the range of 0.010-0.20 mg x mL(-1). The detection limit of the method was 5 microg x mL(-1). The recoveries were 82.47%-98.18% with the relative standard deviation (RSD) of 1.5%-4.9%. The proposed method is sensitive, accurate and rapid with the sampling frequency of 10 samples-h(-1) and suitable for the determination of OPC in the health food.

Objective: To compare the efficacy and safety of endoscopic laser lithotripsy (LL) and endoscopic pneumatic lithotripsy (PL) for ureteral stones., Methods: We retrospectively analyzed the clinical data of 415 patients with ureteral calculi treated with endoscopic laser lithotripsy (n = 214 ) and pnumatic lithotripsy (n = 201 )., Results: The overall successful fragmentation rate of all ureteral stones in a single session of the LL group was higher than that of the PL group (95% vs. 69%, P < 0.01). The average stonefree time of the LL group was shorter than that of the PL group (18 days vs. 31 days, P < 0.01). No complications such as perforation during the operation were observed in the LL group whereas 3 perforations occurred in the PL group., Conclusion: LL has its advantage over PL in its better clinical effect for the stone fragmentation and low complication rate and is an effective and safe treatment for ureteral stones.

Objective: To investigate the molecular pathology in families with long QT syndrome (LQTS) including Jervell-Longe-Nielsen syndrome (JLNS) and Romano-ward syndrome (RWS) and Brugada syndrome (BS) in Chinese population., Methods: Polymerase chain reaction and DNA sequencing were used to screen for KCNQ1, KCNH2, KCNE1, and SCN5A mutation., Results: We identified a novel mutation N1774S in the SCN5A gene of the BS family, a novel mutation G314S in a RWS family which had also been found in Europe, North America, and Japan, and a single nucleotide polymorphisms (SNPs) G643S in the KCNQ1 of the JLNS family. In this JLNS family, another heterozygous novel mutation in exon 2a was found in KCNQ1 of the patients., Conclusion: New mutations were found in our experiment, which expand the spectrum of KCNQ1 and SCN5A mutations that cause LQTS and BS.

Aim: 1) To evaluate the effect of prostaglandin E2 (PGE2) on the regulation of vascular endothelial growth factor (VEGF) expression in gastric MKN28 cells, and 2) to investigate the role of the epidermal growth factor receptor (EGFR) signal transduction pathway in any effect exerted by PGE2 on VEGF expression., Methods: MKN28 cells were incubated with the vehicle (control) or with PGE2 in the presence or absence of AG1478, a selective inhibitor of EGFR tyrosine kinase, or PD098059, a selective inhibitor of the kinase responsible for ERK2 phosphorylation (mitogen-activated protein (MAP)/extracellular signal-regulated kinase (ERK)). Real-time quantitative polymerase chain reaction and Western blot analysis were used to evaluate VEGF mRNA and protein expression. The activity of EGFR and ERK2 was measured by Western blot analysis., Results: PGE2 significantly up-regulated VEGF mRNA and protein expression and increased the activation of EGFR and ERK2. Incubation of MKN28 cells with AG1478 significantly reduced PGE2-induced EGFR activity, ERK2 activity, and VEGF mRNA and protein expression. Meanwhile, incubation of MKN28 with PD098059 reduced PGE2-induced ERK2 activity and VEGF mRNA and protein expression, but had no effect on EGFR activity., Conclusion: Our data suggested that PGE2 up-regulates VEGF expression in gastric cancer cells via transactivation of EGFR-MAPK signaling pathways, which may be mechanisms underlying the contribution of COX-2 to tumor angiogenesis in gastric cancer.

It is reported that the Rhodamine 6G fluorescence in water solution changes with different concentration SDS excited by a 532 nm laser. A small amount of SDS in Rhodamine 6G water solution will reduce the intensity of fluorescence. More SDS enhances R6G fluorescence. The enhancement ratio can reach up to 3.1 when SDS concentration is 6 x 10(-2) mol L(-1) in 5 x 10(-5) mol L(-1) R6G water solution. Dye laser threshold decreases when SDS concentration is 2 x 10(-2) mol x L(-1) in 1 x 10(-4) mol x L(-1) R6G water solution. The physical mechanism of the phenomena is analyzed by the absorption spectra of different concentration R6G water solutions and the fluorescence spectra of R6G water solutions adulterated with different concentration SDS.

Objective: To identify the mutation of a Chinese family with inherited long QT syndrome(LQTS)., Methods: The disease-causing gene was tentatively determined in light of the clinical manifestations and electrophysiological properties, and then polymerase chain reaction and DNA sequencing were used for screening and identifying mutation., Results: A missense mutation G940A(G314S) in the KCNQ1 gene was identified, which was the 'hot spot' of long QT syndrome mutation., Conclusion: The mutation that is involved with long QT syndrome in Chinese patients is the same as that in the European, American and Japanese patients.

In this paper, the authors reported a new method for measuring transient time-dependent spectra using multi-fiber delay coupler and optical multi-channel analysis technique. The fiber delay coupler consists of five fibers with different lengths and its time-delay between two fibers is determined by the different fiber lengths. The light beams that came from the fiber coupler were coupled by a lens system into the slit of a spectrograph and formed a spectral pattern on the focus plane of the spectrometer. A two-dimensional CCD detector converted the optical pattern into an electrical pattern, and then the transient time-dependent spectra were obtained by a personal computer. The experimental setup was constructed, and the transient time-dependent spectra were measured for KCl crystal shocked by a high speed pill. The time resolution was 20 ns, and the spectral intensity resolution reached 1/18 bit.

Tremella aurantia (TA) has been traditionally used as food and crude medicine in Chinese society. The polysaccharide isolated from the fruiting bodies of TA exhibits significant hypoglycemic activity in diabetic mouse models of insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NIDDM). Diabetes will cause sexual dysfunction in patients. In the present study, we examined if the treatment of TA on IDDM and NIDDM rats will restore steroidogenesis and then the reproductive function. The fruiting bodies (FB), mycelium (TM) and polysaccharide (GX) of TA were fed to the IDDM and NIDDM rats, and testosterone and corticosterone levels in plasma, the weight of steroidogenic organs, and the expression of steroidogenic acute regulatory (StAR) protein and P450scc enzyme were determined. Plasma testosterone productions were significantly suppressed with the feeding of FB or TM in normal rat (p < 0.05). Testosterone productions were also significantly suppressed in IDDM diabetes rats (p <0.05), and FB or TM could not restore the inhibitory effects (p > 0.05). There was no significant difference of the testosterone production between normal and NIDDM rats (p > 0.05). In plasma corticosterone production, there were no differences among control, FB- or TM-fed normal rats (p > 0.05). Corticosterone levels were reduced in IDDM rats compared to control, and FB or TM could restore its level. Corticosterone levels were induced in NIDDM rats compared to control (p <0.05), but FB, TM or GX significantly brought the corticosterone back (p < 0.05) to the control levels. Considering steroidogenic organs, IDDM rats with or without TA treatments had heavier testis and adrenal glands, but not epididymis, than normal rats with or without TA treatments. There were no effects of TA on the weight of steroidogenic organs among normal and NIDDM rats. However, GX feeding in NIDDM rat had lesser testis weight compared to NIDDM rats. The expression of StAR protein and P450scc enzyme were not different among groups in IDDM and NIDDM rats. Plasma testosterone productions were suppressed in normal rats with the feeding of TA (FB and TM). IDDM rats did have lower testosterone, but not in NIDDM, and FB or TM could not restore the inhibitory effects. The induction of IDDM or NIDDM rats did affect steroidogenesis and steroidogenic organ weights, and the feeding of TA had different effects on steroidogenesis in different types of diabetic rats.

A new framework for consciousness is introduced based upon traditional artificial neural network models. This framework reflects explicit connections between two parts of the brain: one global working memory and distributed modular cerebral networks relating to specific brain functions. Accordingly this framework is composed of three layers, physical mnemonic layer and abstract thinking layer, which cooperate together through a recognition layer to accomplish information storage and cognition using algorithms of how these interactions contribute to consciousness: (1) the reception process whereby cerebral subsystems group distributed signals into coherent object patterns; (2) the partial recognition process whereby patterns from particular subsystems are compared or stored as knowledge; and (3) the resonant learning process whereby global workspace stably adjusts its structure to adapt to patterns' changes. Using this framework, various sorts of human actions can be explained, leading to a general approach for analyzing brain functions.

This paper discusses the existence and uniqueness of the generalized solution in the sense of Colombeau to the Benjamin-Ono (B-O) equation and the relationship between the new generalized solution and the classical solution.

Objective: Human selenoprotein P (HSelP) is unique protein that contains 10 selenocysteines encoded by 10 inframe UGA, which typically function as stop codon. The function of HSelP remains unclear, in part due to the inability to express it by gene recombinant technique. This study is to investigate expression and purification of recombinant HSelP in prokaryotic expression system, and its activity to induce apoptosis in vitro., Methods: The shorter HSelP isoform was cloned. After the selenocysteine (SeCys) at 40th position from N terminus of the HSelP shorter isoform was mutated into cysteine by PCR, it was expressed in E. coli. The expressed product was purified with DEAE column and identified by Western blot. Subsequently, its function on induction of mitochondrial apoptotic activity was studied., Results: The mutant HSelP shorter isoform expressed in prokaryotic system was purified by DEAE column to 90% homogeneity. The purified product, HSelP280m, induced the opening of mitochondrial permeability transition pore (PTP) and decreased the transmembrane potential in a dose-dependent manner. These events could be abolished by PTP specific inhibitors., Conclusion: HSelP280m can induce the opening of mitochondrial PTP, which provides a basis for investigating the structure and function of recombinant HSelP.

During early embryonic cycles, the time required for mitotic spindle assembly must match the autonomous cell cycle oscillations because a lack of coordination between these two processes will result in chromosome segregation errors. Members of the widely conserved BimC kinesin family are essential for spindle formation in all eukaryotes, and complete loss of BimC function results in monopolar spindles that have two spindle poles that are not separated. However, the precise roles of BimC motor activity in the spindle assembly process are not known. To examine the contribution of BimC kinesin's motor activity to spindle assembly, we generated and characterized mutants of Eg5, a vertebrate BimC kinesin, with reduced in vitro microtubule-gliding velocities. In Xenopus egg extracts, we replaced endogenous Eg5 with recombinant wild-type or mutant motor proteins. By using centrosome-dependent and centrosome-independent spindle assembly assays, we found that mechanisms that determine spindle size and shape were robust to approximately 6-fold reductions in Eg5 motility. However, the spindle assembly process was slower when Eg5 motor function was impaired. This role of Eg5 was independent of its contribution to centrosome separation. We provide evidence that Eg5 is a rate-limiting component of the cellular machinery that drives spindle assembly in vertebrates.

To study PCR site-directed mutagenesis of long QT syndrome KCNQ1 gene in vitro. The site-directed mutagenesis of LQTS gene KCNQ1 was made by PCR. Two sets of primers were designed according to the sequence of KCNQ1 cDNA, and mismatch was introduced into primers. Mutagenesis was performed in a three-step PCR. The amplified fragments from the third PCR which contained the mutation site were subcloned into the T-vecor PCR2.1. Then the fragments containing the mutation site was obtained from PCR2.1 with restriction enzyme digestion and was inserted into the same restriction site of pIRES2-EGFP-KCNQ1. With Effectene Transfection Reagent, pIRES(2)-EGFP-KCNQ1 was transfected into HEK293 cell. The sequencing analysis showed that the mutation site was correct. Mutation from T to C in 934 site of KCNQ1 cDNA was found. Under the fluorescence microscope, the green fluorescence was spread in the transfected HEK293 cell, meaning the pIRES(2)-EGFP-KCNQ1 containing the mutation site was expressed correctly.

The efficacy of angiotensin converting enzyme (ACE) inhibitors in treatment of renovascular disease has been controversial. It has been reported that patients with incidental atherosclerotic renal artery stenosis (ARAS) are sometimes treated with ACE inhibitors before being considered for renal revascularization. This study was designed to describe the frequency and the characteristics of patients with incidental ARAS, and to examine the frequency of ACE inhibitor usage in such patients. We studied a cohort of consecutive patients undergoing abdominal aortography at the time of cardiac catheterization. Patients were stratified and compared based on the presence and severity of ARAS. ARAS (> or =50%) was present in 146 (17.0%) of 859 evaluable patients. Factors independently related to the presence of ARAS were age (odds ratio (OR)=1.07, p < 0.001), severity of coronary artery disease (OR=2.13, p < 0.001) and peripheral vascular disease (OR=1.79, p = 0.021). Among all patients with ARAS, the percentage of ACE inhibitor usage was 74.7% (109/146). Among patients with severe ARAS, moderate ARAS, mild ARAS, insignificant ARAS and normal renal arteries, the percentage of ACE inhibitor usage was 85.7% (95% confidence interval (CI): 69-100%), 82.9% (95% CI: 71-95%), 68.5% (95% CI: 59-78%), 68.6% (95% CI: 55-82%) and 53.9% (95% CI: 50-58%), respectively (contingency coefficient=0.17, p < 0.001). In patients with severe ARAS, ACE inhibitor use, calcium channel blocker use and diuretic use were shown to correlate significantly with serum creatinine levels after controlling for potential confounding factors. In this study, ACE inhibitors were used commonly in patients with incidental ARAS; the frequency of ACE inhibitor use correlated with the severity of ARAS.

A new kind of instrument and method for simultaneous and real-time collection of multi-object spectra by using multi-fiber coupling and Optical Multi-channel Analysis (OMA) was reported. The spectral signals of one object, three objects and five objects were collected successfully in experiments. The spatial resolution of 0.5 mm and detected spectral range of 200-1100 nm were reached, and at most twenty objects could be collected on the image plane of 1 cm2 area with OMA 4 system. The design of the coupling lens's light path was discussed to guarantee enough light energy to enter the instrument and little effect on the resolution of spectral instrument. The instrument can be applied widely to the areas of supervising and controlling the quality of light beam in transmission and in the time-resolved and spatial-resolved spectral measurement.

Objective: To explore the relationship of occupational injuries with social and economic factors in chemical industry during 2000.01 - 2001.12., Method: 1:2 paired case-control study, univariable logistic regression analysis, principal component analysis, and multiple logistic regression analysis were used in this study., Results: Univariable analysis showed that occupational injuries had significant relationship with age, sex, education, employment pattern, technology, workplace, work changing, wage, family income, enterprise scale, enterprise proprietorship, projective device, operation rules, and training rules of work safety. The extracted four principal components (PC(1), PC(2), PC(3) and PC(4), ranked by contribution) gave good expressions to the initial 11 variables. The cumulative proportion of the four principal components reached 77.36%. PC(1) was the indicative factor of occupational injuries, which represented 46.69% information of initial variables. PC(2) was the kinetic factor of occupational injuries. PC(3) was the stable factor of occupational injuries. PC(4) was the sex factor of occupational injuries. The results of multiple conditional logistic regression analysis showed that occupational injuries had statistically significant relationship with PC(1) and PC(2). Among the initial variables, sex, employment pattern, income, scale of enterprise, and property of enterprise were more prominent., Conclusion: Occupational injuries are related with multiple social and economic factors, which often interact on each other. The prevention and control of occupational injuries should require a comprehensive approach, including training and education of work safety, improving workers' consciousness of self-protection, and enhancing proprietors' consciousness of work safety.

The incident pulse intensity wave will cause the induced fluorescence wave, which will lead to the obvious error of experiment results. A double-light-beam correlation method between the incident laser and the fluorescence was used to decrease such error efficiently in the experiments. A good correlation factor over 0.9 was obtained by the method in the experiment of R6G fluorescence enhanced by SDS. The method is better than sampling-averaging method when the incident laser intensity has more waves.

Aim: To prepare rabbit antibody against human selenoprotein P (HSelP) by using HSelP expressed in the prokaryotic expression system and identify its specificity., Methods: HSelP gene fragment was expressed in E.Coli via IPTG induction and purified through DEAE and Ni-NTA columns sequentially. The purified HSelP as immunogen was used to immunize rabbit. The specificity of rabbit anti-HSelP antibody was analyzed by Western blot., Results: HSelP protein had been expressed and purified successfully. The polyclonal anti-HSelP antibody could specifically recognize natural HSelP from human serum., Conclusion: Polyclonal anti-HSelP antibody with high specificity and purity has been prepared by using purified HSelP as immunogen, which lays the foundation for further research on the function and distribution of HSelP, as well as developing a simple method for detecting HSelP.

The objective of this study was to quantify the bioavailability of phenanthrene solubilized in surfactants by a mixed phenanthrene-degrading culture isolated from the petroleum contaminated soils. An nonionic surfactant, Tween20, was used. The effects of Tween20 on the microbial degradation of phenanthrene were evaluated depending on the rotary flasks experiments. The results showed that the concentrations of Tween20 above the critical micelle concentration (CMC) could increase the solubility of phenanthrene on great extent and were not toxic to the phenanthrene-degrading bacteria, and that the presence of surfactant micelles did not inhibit the biodegradation of phenanthrene. Phenanthrene solubilized in the micelles of Tween20 in liquid media was bioavailable and degradable by the mixed culture of bacteria.

Objective: To develop a setup of capillary electrophoresis with laser induced fluorescence detector for separating and detecting DNA fragments and protein., Methods: The setup is assembled of He-Ne Laser with 543.5 nm as the excitation light, filter, monochromator, photomultiplier tube and a computer for controlling the system and collecting the electropherograms., Results: The design of the setup and the analytical parameters were optimized. The electropherograms with high Signal-to-Noise were obtained by the laser induced fluorescence detector-capillary electrophoretic setup. For Sulforhdamine B, the detection limit was 10(-9) mol/L, and the linear range from 1 x 10(-6) mol/L to 1 x 10(-9) mol/L. The PCR products, the digested products by Hae III and BstE of mycobacterium DNA, and the human serum albumin were determined by the successful application of this setup., Conclusion: The setup is characterized by simple structure, low cost and high sensitivity. Compared with traditional agarose gel electrophoresis, capillary electrophoresis with laser induced fluorescence detector is more remarkable in resolution and detection time. It can be applied as a more effective means for analysis of DNA fragments and protein.

Phenanthrene was solubilized in two different nonionic surfactants, Tween80 and Triton X-100. The bioavailability of phenanthrene to the bacteria isolated from the petroleum contaminated soils was studied based on the rotary flasks experiments. The results showed that the concentration of nonionic surfactants above the critical micelle concentration (CMC) can increase the solubility of phenanthrene in water and were innoxious to the phenanthrene-degrading bacteria; phenanthrene solubilized in the micelles of Tween80 was bioavailable and biodegradable. The research demonstrated the potential of surfactant-enhanced bioremediation of soils contaminated by hydrophobic organic compounds (HOCs).

Objective: To assess the effects of laparoscopic retroperitoneal lymph node dissection in the treatment of stage I nonseminomatous testicular cancer., Methods: From January 2001 to May 2002, laparoscopic retroperitoneal lymph node dissection was performed on 9 patients with stage I nonseminomatous testicular cancer., Results: The procedure was successful in all patients. The mean operation time was 260 minutes. None of the patients required blood transfusion and had major complications intraoperatively or postoperatively. The average period of hospitalization after the operation was 5.5 days. With a mean following-up of 9 months, retroperitoneal recurrence was not seen., Conclusions: Laparoscopic retroperitoneal lymph node dissection is feasible for stage I nonseminomatous testicular cancer and its procedure is safe, effective and minimally invasive.

The simultaneous determination of copper and iron by catalytic kinetic spectrophotometric method with back propagating artifical neural net (BP-ANN) was studied based on the catalysis of Cu(II) and Fe(III) to the reaction of rhodamine B hydrogen peroxide in the range of pH 3-9. The determination was performed by the construction of pH gradient and a CCD-diode array detector with a microcomputer. The optimum conditions of reaction and flow injection were studied. The determination limits for copper and iron were 0.10 and 0.21 microgram.L-1, respectively. The sampling rate was 10 samples per hour. The proposed method has been used to determine Cu(II) and Fe(III) in certified reference materials and food samples with satisfactory results. The average values of spiked recovery were 93.3%-100.2% with RSD of 0.6%-7.5%.

Background & Objective: Sep15 is a selenium-containing protein identified in 1998. This protein may be involved in cancer etiology and it may have redox function. The objective of this study was to investigate the relationship between the redox function of Sep15 and tumor development., Methods: The full-length DNA sequence of Sep15 was obtained by RT-PCR and then recombined to eukaryotic expression vector pcDNA3.1(+). The BEL-7402- Sep15 cell line, which expressed the high levels of Sep15 by transfecting the cultured hepatocarcinoma cell line BEL-7402 with pcDNA3.1-Sep15 was generated. From morphologic investigation, cell growth curve, clone formation and nude mice tumor growth curve, the relationship between Sep15 and hepatocarcinoma cell line BEL-7402 was determined. Furthermore, the redox reaction of sep15 was detected by MTT assay., Results: There was no distinct effect of transfection of Sep15 gene on BEL-7402-Sep15 cell. The cell survival rate was drastically different between BEL-7402-Sep15 cell and both BEL-7402- pcDNA cell and BEL-7402-Sep15 cell after foreign H2O2 reactive oxygen stress (P<0.05)., Conclusion: Transfecting Sep15 gene did not influence the growth characteristics of BEL-7402 cell line and Sep15 may have redox function.

Background & Objective: It was reported that endostatin could inhibit tumor angiogenesis, then inhibit the growth and metastasis of tumor. However, there was few report about the treatment usage of endostatin. This study was designed to explore the effect of endostatin mediated with recombinant adeno-associated virus(rAAV) on tumor growth and metastasis., Methods: To obtain the endostatin gene complete cDNA by RT-PCR, and clone it into the plasmid pSNAV and package the recombinant rAAV-SS-endostatin; to analyze its anti-tumor effect through animal experiments., Results: In the B16F10 melanoma tumor model, C57BL/6 mice were inoculated with 10(11)TU rAAV-SS-endostatin i.m. injection, which the inhibition rate was 57.1%; In the lung metastatic model, the inhibition rate of metastases was 70.7%., Conclusions: rAAV-SS-endostatin can effectively inhibit tumor growth and metastasis.

A new kind of method of multi-channel acquisition of thermal radiation spectra and temperature calculating was reported in this paper. Employing a tungsten lamp as thermal source, using an optical multi-channel analyzer(OMA) to collect the radiation spectra, the measured temperatures were calculated by using optimum simulation between the measured spectra and Planck black body radiation curves at the determine temperatures according to the minimum errors principle. The results are agreement with standard values, and the errors are near 1%. The advancing of the method is discussed.

Hyperkalemic solution is widely used to protect the myocardium during open-heart surgery or to preserve donor hearts during heart or heart/lung transplants. The inhibitory effects of hvperkalemic solution on the release of endothelium-derived hyperpolarizing factor (EDHF) of coronary arteries following deep hypothermic storage (4 degrees C) has been well studied. However, it has not been established whether potassium channel openers have protective effects on the coronary endothelial function after cold storage. This study was designed to examine this. Porcine coronary artery rings were studied in organ baths. Relaxation in response to the EDHF stimulus A23187 (nonreceptor-mediated stimulus calcium ionophore) in thromboxane A2 mimetic U46619 (30 nmol/L)-induced precontraction after incubation with hyperkalemic solution (20 mmol/L) with nicorandil (10 micromol/L) (either at 37 degrees C in the oxygenated organ chamber or at 4 degrees C in a refrigerator for 6 h) was compared with the control. There was significant difference between hyperkalemia group and hyperkalemia with nicorandil group under normothermia (p = .04). The difference was significant in the same solution between normothermia and hypothermia. After incubation in hyperkalemic solution without or with nicorandil, the A23187-induced relaxation was 32.8% +/- 9.1% and 72.6% +/- 16.9%, respectively (N = 8, p < .01). Potassium channel opener can attenuate the inhibitory effect of hyperkalemic solution on the release of EDHF after cold storage.

The effect of dexamethasone on follicle-stimulating hormone (FSH)-stimulated expression of cholesterol side-chain cleavage (P450scc) enzyme and production of progesterone by ovarian granulosa cells was studied in vitro. Granulosa cells from 3- to 5-mm pig antral follicles were cultured for 48 h in the presence or absence of FSH and/or dexamethasone. Treatment with FSH resulted in a dose-dependent increase in the level of P450scc mRNA that reached a submaximum at 100 ng FSH/ml. This increase was associated with an increase in progesterone production. Treatment of the cells with increasing concentrations (10(-9)-10(-6) M) of dexamethasone for 48 h increased constitutive and potentiated FSH-stimulated P450scc mRNA levels and progesterone production in a dose-dependent manner. Increasing duration (12-48 h) of treatment with dexamethasone (100 nM) led to a time-dependent increase in basal and FSH-stimulated progesteorne production, achieving statistical significance by 48 and 24 h, respectively. Dexamethasone also increased P450scc mRNA level and progesterone production induced by the adenylate cyclase activator forskolin (10 microM) or a cAMP analog 8-Br-cAMP (1 mM). The effects of dexamethasone on FSH-induced progesterone production were blocked by cotreatment of the cells with glucocorticoid receptor antagonist RU-486. These results demonstrate that dexamethasone potentiates FSH actions on steroidoogenesis in the pig ovary. Possible mechanisms for this potentiation include the ability of dexamethasone to stimulate P450scc gene expression.

Objective: To study the effect of Xanthnotoxol on contractility of isolated rabbit ileum and its relationship with Ca2+., Method: Routine experimental methods for isolated ileum were adopted., Result: Xanthnotoxol (XT) and Verapamil (Ver) inhibited the contraction of isolated rabbit ileum smooth muscle induced by submaximal concentrations of acetylcholine (ACh) and serotonim (5-TH), with a IC50 value (mumol.L-1) of 10.495 +/- 1.521, 0.428 +/- 0.001 and 18.132 +/- 1.627, 0.249 +/- 0.003, respectively. XT and Ver inhibited the contraction induced by Ca2+ after high K+ depolarization and for noncompetitively antagonist CaCl2 cumulative dose-response curve, the pD'2 value was 4.69 +/- 0.03 and 6.35 +/- 0.10, respectively. XT (10 mumol.L-1) and Ver(0.06 mumol.L-1) inhibited the contraction induced by ACh in Ca(2+)-free medium, while XT (100 mumol.L-1) but not Ver(0.6 mumol.L-1) inhibited the extracellar CA(2+)-dependent contraction induced by ACh., Conclusion: XT has a calcium-antagonistic effect which was not similar to that of Ver.

The contents of wall-bound hydroxycinnamic acids and their dimers were compared between elongated and non-elongated cells of suspension-cultured Mentha. Wall-bound peroxidase activity was also investigated. The main hydroxycinnamic acids esterified to these two kinds of cell walls were ferulic and caffeic acids. Eleven dehydrodicaffeic acid isomers and six dehydrodiferulic acid isomers formed through C-C and C-O-C coupling processes, were detected by GC-MS from the extracts released from the walls of non-elongated cells. On the other hand, only four dehydrodicaffeic acid isomers and three dehydrodiferulic acid isomers were found in the walls of elongated cells. Amounts of monomers of ferulic and caffeic acids and their 5,5'-dehydrodimers in non-elongated cell walls were about ten and twenty times higher, respectively, than those in the elongated cell walls. There was a close correlation between the amount of 5,5'-dehydrodimers and activity of wall-bound peroxidase in non-elongated and elongated cells. The level of 5,5'-dehydrodimers accumulated at a higher rate than monomers in non-elongated cell walls. These results suggest that the dimerization of ester-linked ferulic and caffeic acids by peroxidase and the increase in amounts of their 5,5'-dehydrodimers are important factors in the cessation of cell elongation in Mentha suspension culture.

The meiotic division in oocytes is arrested in the G2 phase of the cell cycle. Resumption of meiosis, also known as oocyte maturation, entails a G2 to M transition. At the G2-M boundary, maturation promoting factor (MPF) activation is usually induced via several ways, including tyrosine dephosphorylation of p34(cdc2) and synthesis of cyclin B according to cell type and species. Previous studies in our laboratory demonstrated that glucocorticoids directly inhibit the meiotic maturation of pig oocytes in vitro. The aim of this study was therefore to investigate the influence of glucocorticoids on the expression of p34(cdc2) and cyclin B1 in resumption of meiosis of pig oocytes. We detected the relative levels and association of p34(cdc2) and cyclin B1. Isolated cumulus-enclosed oocytes were cultured in Waymouth MB752/1 medium supplemented with sodium pyruvate (50 microgram/ml), LH (0.5 microgram/ml), FSH (0.5 microgram/ml), and estradiol-17beta (1 microgram/ml) in the presence or absence of dexamethasone (DEX) for 24 hr; they then were cultured without hormonal supplements in the presence or absence of DEX for an additional 24 hr. We found that cyclin B1, as well as p34(cdc2), was already present in fully grown G2-arrested pig oocytes when removed from the follicle. In these oocytes, cyclin B1 and p34(cdc2) were already associated in complex. Treatment with DEX at concentrations of 1 microgram/ml or above decreased the level of cyclin B1, but had no effect on the level of p34(cdc2). The exposure of oocytes to DEX also decreased the amount of complexed p34(cdc2)-cyclin B1. These findings suggest that the inhibitory action of DEX on meiotic maturation could be due, at least in part, to the reduced amount of p34(cdc2)-cyclin B1 complex., (Copyright 2000 Wiley-Liss, Inc.)

Dehydrodicaffeic acid derivatives were found in the cell walls of suspension-cultured cells of Mentha. Using gas chromatography/mass spectrometry (GC-MS) in a single ion chromatography at m/z 790 and m/z 718, eleven peaks of trimethylsilylated dehydrodimers of caffeic acid were detected in the extracts from the cell walls of suspension-cultured cells of Mentha using sodium hydroxide. The result suggests that dehydrodicaffeates are formed in the cell walls from two molecules of caffeate, probably formed through C-C, and C-O-C coupling processes.

Pestalotia heterocornis was isolated from soil collected in yew forest and was shown to produce paclitaxel in semisynthetic liquid media. The presence of paclitaxel in the fungal extract was confirmed by FAB mass spectrometry and NMR spectroscopy. The maximum yield of paclitaxel was 31 &mgr;g per liter. Optimal paclitaxel production occurred after 5-7 days in a 20-liter scale fermentation at 23 degrees C. These results indicate that P. heterocornis is an excellent candidate for consideration in fermentation technology. Copyright 1999 John Wiley & Sons, Inc.

Two distinct types of cell growth of suspension-cultured Mentha were formed when the cells maintained in the medium containing 1000 micrograms l-1 2,4-D were subcultured into different 2,4-D concentrations. Few cell elongation of Mentha (average cell length: 34-40 microns) was observed after division in the medium containing 1-200 micrograms l-1 2,4-D; and significant cell elongation (average cell length: 95-130 microns) was observed after cell division in the medium containing 500-2000 micrograms l-1 2,4-D. A close correlation between culture medium and water content in the cells indicated that 2,4-D promoted cell elongation by water uptake. Amounts of phenolic acid in cell walls were much higher in unelongated cell walls than in elongated ones during the cultivation, and there was a close correlation between the amounts and the level of PAL activity in elongated and unelongated cells. However, there was no significant difference in cell wall components and its neutral sugar composition between elongated and unelongated cells.

The aim of this study was to assess the possible role of glucocorticoids in the maturation of pig oocytes and their subsequent fertilizing capacity in vitro. Pig cumulus-enclosed oocytes collected from prepubertal gilts were cultured in Waymouth MB752/1 medium supplemented with sodium pyruvate (50 microg/ml), LH (0.5 microg/ml), FSH (0.5 microg/ml), and estradiol-17beta (1 microg/ml) in the presence or absence of cortisol or dexamethasone (DEX) for 24 h; they then were cultured without hormonal supplements in the presence or absence of cortisol or DEX for an additional 16-24 h. Treatment of cumulus-enclosed or denuded oocytes with increasing concentrations of cortisol or DEX for 48 h resulted in a dose-response inhibition of germinal vesicle breakdown (GVB). Increasing duration (12-48 h) of treatment with DEX (10 microg/ml) led to a time-dependent inhibition of GVB, which achieved statistical significance by 12 h. The addition of DEX (10 microg/ml) to maturation medium immediately after culture or at 12 h, 24 h, or 36 h after culture also decreased the percentage of oocytes with GVB. When oocytes were exposed to DEX for 48 h, the maturation rate was reduced. The degree of this reduction was dependent on DEX, and a concentration of DEX higher than 0.1 microg/ml was needed. The inhibitory effect of DEX on the maturation of oocytes was prevented by the glucocorticoid receptor antagonist RU-486. Exposure of oocytes to DEX for 40 h did not prevent sperm penetration, affect the incidence of polyspermy, or decrease the ability of oocytes to form a male pronucleus. The intracellular concentration of glutathione (GSH) in cumulus-enclosed oocytes was 4.4 mM per oocyte. Exposure of oocytes to DEX (0.01-10 microg/ml) had no effect on GSH concentration. These results demonstrate that glucocorticoids directly inhibit the meiotic but not cytoplasmic maturation of pig oocytes in vitro. This inhibitory effect is not mediated through a decrease in the level of intracellular GSH.

This study examines the effects of cyclophosphamide, a widely used anti-cancer agent, on the maturation of pig oocytes and on their subsequent fertilizing capacity in vitro. Pig cumulus-oocyte complexes collected from prepubertal gilts were cultured in Waymouth MB 752/1 medium supplemented with sodium pyruvate (50 micrograms/ml), luteinizing hormone (0.5 microgram/ml), follicle-stimulating hormone (0.5 microgram/ml), and 17 beta-estradiol (1 microgram/ml) in the presence or absence of cyclophosphamide for 24 hr; they then were cultured without hormonal supplements in the presence or absence of cyclophosphamide for an additional 16-24 hr. The breakdown of germinal vesicle (GVBD) and changes in glutathione (GSH) content before in vitro fertilization were assessed. Oocytes containing one polar body and a metaphase plate were regarded as matured. Cytoplasmic maturation as determined by male pronuclear formation following fertilization in vitro was also examined. Treatment of oocytes with increasing concentrations (1-1000 micrograms/ml) of cyclophosphamide for 48 hr resulted in a dose-response inhibition of the rate of maturation, but had no effect on GVBD. Increasing duration (12-48 hr) of treatment with cyclophosphamide (100 micrograms/ml) led to a time-dependent inhibition of nuclear maturation, achieving statistical significance by 24 hr. The addition of cyclophosphamide (100 micrograms/ml) to maturation medium immediately after culture, 12 hr or 24 hr after culture also decreased the percentage of oocytes matured during a 48-h culture period. Exposure of oocytes to cyclophosphamide (100 micrograms/ml) for 40 hr did not prevent sperm penetration, not affect the incidence of polyspermy, or decrease the ability of oocytes to form a male pronucleus at 8 hr after insemination. The concentration of GSH, an important factor for male pronuclear formation, in pig oocytes was determined by an enzymatic cycling assay. The concentration found was 8.15 +/- 1.19 mM per oocyte. Exposure of oocytes to cyclophosphamide (100 micrograms/ml) had no effect on GSH concentration. These results demonstrate that cyclophosphamide directly inhibits the meiotic but not cytoplasmic maturation of pig oocytes in vitro. This inhibitory effect, apparently, is not mediated through a decrease in the level of intracellular glutathione.

The 5' untranslated sequence (5' UTS) of platelet-derived growth factor B (PDGF-B/c-sis) mRNA is highly preserved through evolution, and inhibits translation of downstream coding sequences. In this study, using Northern analysis we identified two PDGF-B/c-sis mRNAs (3.5 kb and 2.6 kb) expressed in normal developing rat brain. In contrast to the constitutive expression of 3.5 kb mRNA, the expression of 2.6 kb mRNA increased markedly in accordance with those stages of brain development at which we had previously demonstrated an increased immunoreactivity for PDGF-B/c-SIS in neurons (Sasahara et al., 1992). By PCR cloning and the RNase protection assay, we determined the complete sequence of rat PDGF-B/c-sis, and found that the 2.6 kb transcript was a form of the 3.5 kb message truncated at the 5' end, and that the predominant 2.6 kb mRNA commenced 15 nt upstream of the signal peptide. Accordingly, it is suggested that the truncation of 5' UTS contributes to the expression of PDGF-B/c-SIS protein in the CNS. Lack of translational inhibitory 5' UTS of PDGF-B/c-sis transcript and resultant efficient protein translation have been reported in only a few transformed cells and cultured umbilical vein endothelial cells. We have extended this knowledge to the developing rat brain, and suggest that a similar mechanism could operate widely in non-transformed tissue in vivo.

We investigated whether there is any difference in optic nerve head topography between primary open-angle glaucoma (POAG) and normal-tension glaucoma (NTG) in Koreans. Twenty-three eyes of 23 POAG and 20 eyes of 20 NTG Korean patients closely matched for mean deviation of automated static threshold perimetry (Humphrey Field Analyzer, program C30-2) were enrolled. The patients' mean age was 51.5 +/- 11.7 years in POAG and 55.1 +/- 10.6 years in NTG; the average mean deviation was -5.47 +/- 1.84 dB in POAG and -6.45 +/- 3.69 dB in NTG. Mean age and mean deviation were not statistically different between the two groups. Heidelberg Retina Tomograph was used to obtain topographic measurements of disc area, rim area, cup volume, cup-to-disc area ratio, mean cup depth, maximum cup depth, and cup shape measure. Disc area and cup volume were significantly larger in NTG than in POAG (p < 0.01, p = 0.03, respectively, Wilcoxon rank sum test). Other variables were not significantly different between the two groups. There was significant negative correlation between cup volume and rim area only in POAG (r = -0.725, p < 0.01, Spearman correlation analysis). These results suggest that the larger cup volume in NTG than in POAG may be influenced rather by larger optic disc in NTG not by smaller neuroretinal rim.

Misalignment between the two elements of the CMT1A-REP binary repeat on chromosome 17p11.2-p12 causes two inherited peripheral neuropathies, Charcot-Marie-Tooth type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies. This binary repeat contains repetitive DNA elements that include LINES, SINES, medium reiteration frequency repeats, and a transposon-like element. The COX10 gene has been mapped 10 kb centromeric to the distal CMT1A-REP element, and a portion of this gene is present in both the proximal and the distal CMT1A-REP elements. We report the isolation and characterization of a novel cDNA (C170RF1), which maps centromeric to and partially within the proximal CMT1A-REP element. Part of C170RF1 is transcribed from the opposite strand of the COX10 partial gene duplication present in the proximal CMT1A-REP element. This finding shows that C170RF1 and COX10 are being transcribed from opposite strands of identical DNA sequences that are separated by 1.5 Mb in the genome. RT-PCR analysis showed the proximal transcript was expressed in skeletal muscle. Sequence analysis identified an open reading frame encoding a 199-amino-acid protein. Zoo blot analysis showed that the transcript is conserved in nonhuman primates. The presence of a binary repeat contributes to the instability of this region of chromosome 17, yet two CMT1A-REP elements are present in the chimpanzee and all human populations. The presence of expressed sequences in both elements of the CMT1A-REP binary repeat could explain the maintenance of this repeat in humans.