Search

Objective To summarize the characteristics of cognitive function and sleep structure in patients with idiopathic rapid eye movement sleep behavior disorder (iRBD) and to explore the correlation between them. Methods Total 73 patients with iRBD who visited Fujian Medical University Union Hospital from August 2018 to August 2021 were included, all of whom underwent neuropsychological tests and polysomnography monitoring. Pearson and partial correlation analyses were used to explore correlation between cognitive function and sleep structure parameters. Results 1) Neuropsychological tests: iRBD patients had lower Mini ⁃ Mental State Examination (MMSE; t = ⁃ 3.703, P = 0.000), Montreal Cognitive Assessment (MoCA; t = ⁃ 4.811, P = 0.000), Verbal Fluency Test (VFT)⁃number of correct words (t = ⁃ 2.171, P = 0.032) than control group. Trail Making Test (TMT)⁃A (t = 2.500, P = 0.014), TMT⁃B (t = 2.430, P = 0.016) and Stroop Color Word Test (SCWT)⁃A (t = 2.507, P = 0.013), SCWT⁃B (t = 15.042, P = 0.000), SCWT⁃ C (t = 27.228, P = 0.000) completion times were longer than control group. 2) Polysomnography monitoring: total sleep time (t = 2.699, P = 0.008), sleep efficiency (Z = ⁃ 2.103, P = 0.035), number of sleep stage transitions (t = 3.965, P = 0.000), proportion of non⁃rapid eye movement sleep stage 1 (N1 stage) to total sleep time (t = 2.887, P = 0.005), periodic limb movement of sleep index (PLMSI; t = ⁃ 2.917, P = 0.004), periodic limb movement⁃related arousal index (Z = ⁃ 2.291, P = 0.022), and total arousal index (Z = ⁃ 2.609, P = 0.009) were higher than those in control group, and the time of wakefulness after sleep was less than those in control group (t = ⁃ 2.230, P = 0.027). 3) Correlation between cognitive function and sleep structure parameters: MoCA score was negatively correlated with the percentage of N1 stage (r = ⁃ 0.184, P = 0.035), and positively correlated with the percentage of N2 stage (r = 0.173, P = 0.049); SCWT⁃B completion time was negatively correlated with the number of awakenings (r = ⁃ 0.186, P = 0.033); SCWT⁃C completion time was positively correlated with the number of sleep phase transitions (r = 0.212, P = 0.015) and the percentage of N1 stage (r = 0.181, P = 0.039); VFT⁃number of correct words was negatively correlated with the periodic limb movement of wake index (PLMWI; r = ⁃ 0.216, P = 0.018); the number of correct China⁃Rey Auditory Verbal Learning Test (C⁃RAVLT) immediate recall was positively correlated with the rapid eye movement (REM) sleep latency (r = 0.183, P = 0.045); the number of correct C⁃RAVLT delay recall was negatively correlated with the PLMWI (r = ⁃ 0.196, P = 0.032) and PLMSI (r = ⁃ 0.180, P = 0.049); the Clock Drawing Test was positively correlated with sleep latency (r = 0.192, P = 0.035) and REM sleep latency (r = 0.199, P = 0.029), and negatively correlated with the percentage of REM sleep (r = ⁃ 0.189, P = 0.038). The Beck Depression Inventory (BDI) score was positively correlated with total sleep time (r = 0.347, P = 0.000), sleep efficiency (r = 0.319, P = 0.000) and the percentage of REM sleep (r = 0.204, P = 0.026), and negatively correlated with the time of wakefulness after sleep (r = ⁃ 0.280, P = 0.002) and the percentage of N1 stage (r = ⁃ 0.299, P = 0.001). Conclusions Patients with iRBD have cognitive dysfunction mainly manifested by decreased attention, executive function and language function, and sleep structural disorders may affect the cognitive function of such patients.

BACKGROUND Eco-functional areas play an important role in soil and water conservation and biodiversity conservation. The Wushui River is a watershed located in the Nanling ecological function area. However, study on the content and potential ecological risk of heavy metals in the soil of the upstream area of the watershed is scarce. OBJECTIVES To investigate the degree of heavy metals pollution and the potential ecological risk of soils in the upper reaches of the Wushui River. METHODS The soil samples of five land utilization types in the upper reaches of the Wushui River Basin, including mining, industrial, and cultivated land and forest, were collected to analyze the contents of seven heavy metals Cd, As, Cu, Hg, Ni, Pb and Zn in the soil. The Nemerow comprehensive pollution index was used to evaluate the degree of heavy metal pollution, and the Hakanson potential ecological risk index was used to evaluate the potential ecological risk of heavy metals in soil. Principal component analysis was used to investigate the source of heavy metal pollution. RESULTS The results show that the average concentrations of Cd, As, Cu, Hg, Ni, Pb and Zn in the upper reaches of the Wushui River were 1.28, 72.44, 54.62, 0.27, 68.32, 72.29 and 158.42mg/kg, respectively, which higher than the soil background values. The contents of the heavy metals, except mercury in mining land, were higher than those in other types of soils. The single factor pollution index of heavy metals in the mean state show that Cd and As were heavy pollution elements with factors of 5.07 and 3.25, respectively. In mining land, the Cd single factor pollution index reached 13.59. Heavy metal pollution in mining land was the most serious, followed by industrial land. Forest was in a safe state. Potential ecological hazard index evaluation results showed that Cd reached a very strong ecological hazard and As was a strong ecological hazard in mining land. Mining land and industrial land reached a strong ecological hazard, industrial land reached a moderate ecological hazard, and other types of soils were slightly ecologically harmful. The results showed that As, Cd, Cu and Zn came from industrial and mining activities, Ni and Hg mainly came from parent materials, and Pb came from transportation. CONCLUSIONS Heavy metal pollution of soil is relatively serious in the upstream area of the Wushui River Basin. Cd and As are the main risk factors in the area, mainly deriving from mining and industrial activities.

In order to standardize and complete the radiated interference E-field threshold testing in reverberation chambers proposed previously, the influence factors on the measurement accuracy were analyzed in this paper, obtaining the error variation with different test conditions. It is revealed that the electrical size of the equipment under test (EUT) is the main restricted factor to the application range of the proposed method. Decreasing the electrical size of the EUT, an experiment was implemented to study the influence of the electrical size on measurement accuracy. The result shows that the measurement error can be decreased to less than 3 dB when the testing factors are reasonably chosen and the electrical size of the EUT is large enough. However, when the electrical size of the EUT is too small, the measurement uncertainty level may be larger than 3 dB. Accordingly, the proposed method is more proper for the EUTs with large electrical sizes. [ABSTRACT FROM AUTHOR]

The design of objective lens for low-light night vision system with plastic and glass optical elements is introduced. The optical athermalization method is used for temperature change and aberration balancing. The operating wavelength is in the range from 0. 65 µm to 0. 95 µm, the field of view is 40°, and F-number is 1.2. The modulation transfer function (MTF), temperature adaptability and weight of night vision optical system are analyzed and evaluated by the CODE V optical design software. The design result indicates that MTF of all FOVs is more than 0.4 at 40 lines per millimeter in the total temperature range and the weight of all lenses is less than 20 g. The optical system obtains weight reduction and better image quality. [ABSTRACT FROM AUTHOR]

To consider dynamic load while equivalent standard axle load conversion is carried out, two degree of freedom vehicle model is built to calculate power spectral density of dynamic load coefficient, and then distribution of dynamic load coefficient is calculated. Increasing degree of equivalent standard axles is calculated in theory while dynamic load is considered. The results of examples show that equivalent standard axle load shall be increased when dynamic load is considered for various kind of pavement: for asphalt pavement, while design deflection and tensile stress at the bottom of asphalt pavement is taken as index, 1% - 2% shall be increased for expressway and first class highway, 2% - 5% shall be increased for other classes highway; while tensile stress at the bottom of semi-rigid base is taken as checking index, 3% - 6% shall be increased for expressway and first class highway, 7% - 17% shall be increased for other classes highway; for cement concrete pavements, 15% - 25% shall be increased for expressway and first class highway, 35% -80% shall be increased for other classes highway. [ABSTRACT FROM AUTHOR]

Karst water, which provides approximately 25% of the world's drinking water, is especially vulnerable to anthropogenic pollutants. To determine the variations between high and low flow periods and the sources of dissolved sulfate (SO 4 2- ) in small karst basins, SO 4 2- concentrations, stable sulfur and oxygen isotopes ( δ 34 S-SO 4 and δ 18 O-SO 4 ), and oxygen isotopes of water ( δ 18 O-H 2 O) were investigated in surface and groundwaters, during the high and low flow seasons, within the Babu subterranean stream basin. Analysis showed that: ① the water samples that were directly impacted by acid mine drainage exhibited high SO 4 2- concentrations (≥250 mg·L -1 ) and significant seasonal variation, while the seasonal variation of non-AMD-impacted water with low SO 4 2- concentrations was not significant. ② During the high flow season, the mean δ 34 S-SO 4 and δ 18 O-SO 4 values of surface water were -10.5‰ and 4.7‰, respectively, and -11.5‰ and 1.3‰ during the low flow period; the mean values of δ 34 S-SO 4 and δ 18 O-SO 4 in groundwater samples were -2.9‰ and 7.1‰ during the high flow period, respectively, and -3.2‰ and 6.2‰ during the low flow period. Both surface and groundwater samples exhibited higher δ 34 S-SO 4 and δ 18 O-SO 4 values during the high flow period than during the low flow period. ③ The values of δ 34 S-SO 4 in the surface and groundwater samples were relatively stable, indicating that the sources of SO 4 2- at specific sampling sites were stable.④ The main sources of SO 4 2- in surface and groundwaters were rain, sulfide, and gypsum, accounting for 13%, 40%, and 47%, respectively, of SO 4 2- in samples taken from the basin outlet during the high flow season, and 18%, 39%, and 43%, respectively, in samples obtained during the low flow season.

Multiple isotopes (C, N, and O) and hydrochemical data were used to trace the sources and fate of nitrate in ground and surface waters of the Babu subterranean stream watershed in Guizhou Province. The origin of nitrate in the water samples was also quantitatively analyzed by the SIAR model. Approximately 38% of the groundwater samples were not drinkable because the nitrate exceeded the drinking water standard, thereby indicating that the groundwater was seriously polluted by nitrate. The ranges of δ 15 N-NO 3 , δ 18 O-NO 3 , and δ 18 O-H 2 O in groundwater were 2.30‰-30.33‰ (mean of 9.68‰), 2.65‰-13.73‰ (mean of 6.64‰), and -8.83‰﹣-7.37‰ (mean of -8.18‰), respectively. Based on the stable isotopic compositions ( δ 15 N-NO 3 , δ 18 O-NO 3 , and δ 18 O-H 2 O), nitrification was the dominant process in the basin. The nitric acid produced by nitrification promoted the dissolution of carbonate rocks, thereby leading to a significantly negative correlation ( P <0.001) between the carbon isotope of dissolved inorganic carbon ( δ 13 C DIC ) and δ 15 N-NO 3 and indicating that δ 13 C DIC , combined with δ 15 N-NO 3 , is effective in exploring the fate of nitrate in karst groundwater. The nitrate in the ground and surface waters mainly originated from soil N, manure and sewage, and ammonium nitrogen fertilizer. The results of the SIAR model showed that the contributions of soil N, manure and sewage, and ammonium nitrogen fertilizer were 36.19%, 33.71%, and 30.10% in groundwater, respectively, and 39.15%, 36.08%, and 24.77% in surface water, respectively. Therefore, it would be more effective to reduce the nitrate recharge flux in groundwater by simultaneously removing nitrate and ammonium nitrogen during wastewater treatment and by adopting scientific fertilization technology in agricultural areas.

Based on the seasonal characteristics of groundwater hydrochemistry and the carbon isotopes ( δ 13 C) of dissolved inorganic carbon (DIC) in the Hongjiadu Basin, Guizhou Province, this paper discusses the natural processes and anthropogenic factors affecting the characteristics of δ 13 C DIC in karst groundwaters under different land use types. The results show that the main sources of DIC in groundwater are carbonate weathering and soil CO 2 . In winter, the δ 13 C DIC values for groundwater ranged from -14.8‰ to -4.1‰ with an average of -10.1‰ and, in summer, ranged from -14.5‰ to -6.3‰ with an average of -10.2‰. Sulfuric acid from sulfide oxidation in coal-bearing strata and acid rain is involved in carbonate weathering, resulting in the enrichment of groundwater with heavy carbon isotopes. Due to the soil CO 2 effect, the δ 13 C DIC values of woodland groundwater experiencing less disturbance from human activities are lower in summer than in winter. The degradation of organic matter input from residential areas is a significant contributor of DIC to groundwater. The average values of δ 13 C DIC in winter and summer were -11.9‰ and -11.6‰, respectively, and the seasonal difference was relatively small in residential areas. During different seasons and for different types of land use, human activities could lead to differences in groundwater δ 13 C DIC values and hydrochemistry. Therefore, δ 13 C DIC can reflect the impact of human activities on karst aquifers, which has important ecological significance.

Chemical fertilizer plays an important role in increasing grain production in agricultural systems but overuse also brings a series of environmental problems, such as eutrophication of surface water, deterioration of soil structure, and the decline of agricultural carrying capacity. At present, research on chemical fertilizer use mainly focuses on utilization efficiency while studies on the spatial characteristics of its use are limited. It is also of great significance for the sustainable development of agriculture in China to fully understand the spatial changes in the use of chemical fertilizers. Based on data of pure chemical fertilizer use, sown area, and grain yield in prefecture-level cities from 2000 to 2015, spatial autocorrelation analysis, cold and hot spot analysis, nuclear density analysis, and standard deviation ellipse analysis were applied. The temporal and spatial variations in total fertilizer use and fertilizer load per unit area in 2000, 2005, 2010, and 2015 were compared. The results showed that:① The use of chemical fertilizer in China increased linearly from 2000 to 2015 but it has been in a relatively high agglomeration state in the region. The pure use of chemical fertilizer experienced a process of increasing regional aggregation from 2000 to 2005, weakening from 2005 to 2010, and increasing from 2010 to 2015. ② From a spatial perspective, the total amount of chemical fertilizer applied between 2000 and 2015 showed an increasing trend, shown by an increase in the number of prefecture-level cities and regions in which the use of chemical fertilizer exceeded the standard. The eastern regions of China, which have experienced rapid economic development, such as Guangdong Province and Fujian Province, are over the critical load of chemical fertilizer more and denser than other prefecture-level cities because the proportional reduction in sown area is greater than the increase in chemical fertilizer use. ③ The movement track of the grain production center in China from 2000 to 2015 was not consistent with the movement track of chemical fertilizer usage. Specifically, the center of grain production moves to the northeast, while the center of chemical fertilizer use moves to the west. This demonstrates that the status of commercial grain production in Northeast China is becoming more and more important, and that the use of chemical fertilizer in the western regions of China is gradually increasing. Here, there is a trend for exceeding the standard, which required further attention.

In recent years, the ecological environment of wetlands has deteriorated under the predatory exploitation of human beings. Nitrate pollution is a major problem faced by wetland ecosystems. In this paper, Huixian karst wetland was selected as a study area to identify the main factors leading to the increase of nitrate in wetland water. The main sources of nitrate pollution in the area were determined by means of 15 N(NO 3 - ) and 18 O(NO 3 - ) isotopes. The contribution rate of all types of pollution sources was determined with the SIAR model. To explore the relationship between the spatial distribution of the nitrate concentration and the groundwater runoff conditions, the 13 C DIC isotopes were used to qualitatively characterize the runoff conditions of the groundwater. The results show that there are three main factors affecting the nitrate concentration in the study area:animal manure and domestic sewage, NO 3 - in chemical fertilizer, and soil nitrogen. The average contribution rates to nitrate pollution are 39.1%, 32.2%, and 28.5% respectively. The 13 C DIC isotope data clarify that a light 13 C DIC reflects better groundwater runoff conditions. The concentration of NO 3 - is generally lower, while the NO 3 - concentration corresponding to heavy 13 C DIC is generally higher. The condition of the groundwater runoff to a certain extent also affects the distribution of the NO 3 - concentration.

Objective: To study whether Exendin-4(Ex-4) could influence oxidative stress in PC12 cells induced by methylglyoxal and its underlying mechanism., Methods: PC12 cells were cultured with methylglyoxal (0,0.25,0.50,0.75,1.0,2.0 mmol/L) for 12~48 h, or PC12 cells were pretreated with Ex-4 (25, 50, 100, 200 nmol/L) for 24 h then incubatedwith methylglyoxal (0.75 mmol/L) for 24 h. MTT assay was used to measure cell viability. Fluorescent probe method was used to detect reactive oxygen species (ROS) expression. Xanthine oxidase method was used to detect superoxide dismutase (SOD)activity. With pretreatment of Exendin-4 (100 nmol/L) for 24 h,the expressions ofP-IκBα, Inhibitor of NF-κB-α IκBα were detected by Western blot after PC12 cells were exposed to methylglyoxal (0.75 mmol/L) for 1 h., Results: Following methylglyoxal administration, cell viability was gradually decreased in a dose-and time-dependent manner. Pretreatment with Ex-4 for 24 hours, cellviability were gradually increased compared with methylglyoxal-alone group. Pretreatment with Ex-4 (100 nmol/L) for 24 hours, ROS expression was reduced by65.30% ( P <0.01) compared with methylglyoxal-alone group, ROS expression in NAC-pretreatment group was reduced by 107.40% ( P <0.01); SOD activity in the Ex-4 pretreatment group was increased by 5.30 U/mg prot ( P <0.01), SOD activity in the NAC pretreatment group was increased by 8.53 U/mg prot ( P <0.01);the ratio of P-IκB-α/IκB-α in the Ex-4 pretreatment group was reduced by 25.50% ( P <0.01), the ratio of P-IκB-α/IκB-α in the NAC pretreatment group was reduced by 35.14% ( P <0.01)., Conclusions: This study demonstrates that Ex-4 can increase the viabilities of PC12 cells and protect PC12 cells from oxidative stress induced by methylglyoxal, the mechanism may involve in suppressing the activation of protein IκB-α.

Objective: To identify the predictors of the positive results of transrectal ultrasound (TRUS)-guided biopsy for prostate cancer., Methods: We performed univariate and multivariate logistic regression analyses on the relevant data on 385 male patients that underwent TRUS-guided biopsy for prostate cancer, including such potential predictors as age, body mass index (BMI), symptoms, results of digital rectal examination (DRE), tPSA, fPSA, free/total PSA ratio (f/tPSA), prostate volume (PV), and PSA density (PSAD) for identification of the risk factors related to the positive rate of biopsy. Then we constructed a scoring system as a tool for predicting prostate cancer in repeat biopsies and determined the sensitivity of the system by calculating the false positive rate using the receiver operating characteristic curve., Results: Among the 385 patients, 139 (36.1%) were diagnosed with prostate cancer. On multivariate analysis, age (P < 0.01), DRE (P < 0.01), tPSA (P < 0.01), fPSA (P < 0.01), f/tPSA (P < 0.01), PV (P < 0.01), and PSAD (P < 0.01) were all significant predictors of prostate cancer. Multivariate logistic regression analysis showed age, tPSA, f/tPSA, PV, and PSAD to be independent predictors, with ORs and 95% CIs of 1.07 (1.05-1.16), 1.05 (1.02-1.15), 0.97 (0.86-0.99), 0.98 (0.87-0.96), and 1.79 (1.48-2.06), respectively. Moreover, patients with the risk score of 3-5 had a significantly higher rate of prostate cancer than those with 0-2 (64% vs 11%, P < 0.001)., Conclusion: The scoring system on the key predictors of prostate cancer can help urologists to identify the men in need of prostatic biopsy.

Objective: Eplets mismatch based on HLAMatchmaker software evaluates the clinical application of kidney transplantation., Methods: In 239 cases of renal transplant,merits of methods of the traditional HLA six antigen matcheing criteria, cross reaction groups standard and Eplets mismatch based on HLAMatchmaker standard were compared respectively., Results: The number of mismatchs with three methods in 239 cases, were grouped according to low-high mismatchs. The results revealed that HLAMatchmaker algorithm could significantly increase the number of low mismatchs group 54 (22.6%), compared with the HIA group 19(7.9%) and CREGs group 32 (13.4%). The comparison was discovered statistical significance among the three groups (P<0.001), so the comparison between each group was., Conclusion: HLAMachmaker of donor-recipients matching, is a more efficient, time-saving and high sensitivity matching solution to allograft renal transplantation.

Objective: To explore the effects of aging on the levels of reproduction-related mRNA genes including Gnrh, KISS1/KISS1r, estrogen receptor-alpha (ERα), estrogen receptor-beta (ERβ) and progesterone receptor (PR) in hypothalamus., Methods: Proestrus and metestrus in young (3-4 months) and middle-aged (10-11 months) female mice and diestrus in senile (18-19 months) female mice were observed. And the levels of related mRNA genes in preoptic area anterior hypothalamus (POA-AH) and medial basal hypothalamus (MBH) were determined by real-time polymerase chain reaction (RT-PCR)., Results: In middle-aged mice on proestrus, the level of Gnrh mRNA in POA-AH (0.896 ± 0.049) was significantly lower than that in young mice (1.228 ± 0.147, P = 0.049). The level of ERα mRNA in POA-AH decreased in young mice on proestrus whereas increased in middle-aged mice (0.432 ± 0.063 vs 0.603 ± 0.018, P = 0.016). The level of ERα mRNA of POA-AH, both in middle-aged mice (0.432 ± 0.063, P = 0.014) and senile mice (0.403 ± 0.145, P = 0.020) on diestrus, were significantly lower than that in young mice. The PR mRNA expression in middle-aged mice on proestrus (1.037 ± 0.037) was markedly lower than that in young mice (1.251 ± 0.081, P = 0.031) . In senile mice, the levels of Gnrh mRNA (1.520 ± 0.146, P = 0.004) and ERβ mRNA (1.572 ± 0.184, P = 0.011) increased in POA-AH compared with that in young mice on metestrus. Aging had no effect upon KISS1 and KISS1r mRNA levels in POA-AH. In contrast, KISS1 mRNA level of MBH in middle-aged (1.663 ± 0.398, P = 0.037) and senile (2.622 ± 0.454, P = 0.014) mice obviously increased compared with the young mice group., Conclusion: Higher levels of ERα mRNA and decreases of PR and Gnrh mRNA in POA-AH in middle-aged mice on proestrus may play an important role in declining reproductive function.

Objective: To explore the impact and related mechanisms of stromal cell-derived factor-1α (SDF-1α) on serum deprivation-induced apoptosis of cardiac stem cells (CSCs)., Methods: CSCs were isolated from adult mouse heart tissue and cultured in vitro. Obtained cells were purified using magnetic-activated cell sorting (MACS) with c-kit magnetic beads. C-kit(+)CSCs were divided into five groups: normal control group, serum deprivation group, serum deprivation+SDF-1α group, serum deprivation+SDF-1α+AMD3100 group, serum deprivation+SDF-1α+LY294002 group. Cell apoptosis was assessed using the DeadEnd Colorimetric TUNEL System and flow cytometry analyses with an Annexin V-FITC Apoptosis Detection Kit. The viability of CSCs was assessed by CCK-8. The protein expression of Bcl-2 and phosphorylated Akt were detected by Western blot. The caspase-3 activity was determined using caspase-3 Colorimetric Assay Kit., Results: After magnetic separation, more than 85% of cardiosphere derived cells were positive for c-kit expression. Compared with the normal control group, the apoptosis rate of serum deprivation group was significantly increased[(27.03 ± 0.80)% vs. (1.51 ± 0.54)%, P < 0.01], which could be significantly reduced by SDF-1α in a concentration dependent manner and peak effect was seen with 100 ng/ml SDF-1α[(10.67 ± 1.06)% vs. (27.03 ± 0.80)%, P < 0.01]. The expressions of p-Akt and Bcl-2 were significantly increased and the activity of caspase-3 was significantly decreased in serum deprivation+SDF-1α group compared to serum deprivation group (P < 0.01). Further more, the expression of p-Akt and Bcl-2 were significantly decreased and the activity of caspase-3 was increased in both serum deprivation+SDF-1α+AMD3100 group and serum deprivation+SDF-1α+LY294002 group compared to serum deprivation+SDF-1α group (P < 0.01)., Conclusions: SDF-1α reduces serum deprivation induced CSCs apoptosis via modulating PI3K/Akt signaling pathway.

To establish a fluorescent quantitative PCR method (FQ-PCR) with TaqMan probe for simultaneous detection of polyomavirus (BKV) and cytomegalovirus (CMV) and to evaluate its clinical application in the renal transplantation recipients. The conservative sequences of BKV and CMV were targeted and amplified by nested PCR technique. The PCR products were cloned into the plasmids pcDNA3. 1(+). The recombinant plasmid containing target sequences of BKV and CMV were constructed as external standards. The TaqMan-based assay was optimized. For evaluating the assay, the sensitivity was determinated by diluted standard (5 X 103-10icopies/mL), and the specificity was verified by negative control and positive control, and the precision was assessed by intra-assay coefficient of variation (ICV) through detecting standard repeatedly (20 times). A total of 480 blood samples of renal transplantation recipients were used to detect BKV and CMV DNA simultaneously with FQ-PCR, and the concentrations of FK506 were measured by ELISA. The association of DNA copy and concentrations of FK506 was analyzed. The cloned target BKV and CMV DNA was confirmed by sequencing and analysis. The sensitivity of the FQ-PCR assay reached 5 X 103 copies/ml in detecting BKV or CMV DNA. Control DNA verified the assay specifically detecting target DNA. The precision of the assay to quantif target DNA copies was acceptable (Intra-assay CV was 3.44% for BKV and 2.23% for CMV; Inter-assay CV was 4. 98% for BKV and 3.76% for CMV;). Of 480 samples, 130 samples (27. 08%) were CMV DNA positive, significantly higher than the BKV DNA positive (13.33%, 64/480, P<0.05). The positive BKV or CMV DNA was found to be associated with high concentrations of FK506 (P<0. 05). In conclusion, the developed real-time PCR assay for detecting both CMV and BKV DNA simultaneously was s high sensitive, precise and time-effectiveand could be applied in the monitoring of the CMV and BKV infection in the renal transplantation recipients.

Objective: We aimed to establish a quantified method for the 17 phthalate acid esters (PAE) in edible vegetable oil by gas chromatography-mass spectrometry (GC-MS) with the pretreatment of acetonitrile extraction and silica/N-(n-propyl)ethylenediamine (silica/PSA) mixed solid phase extraction column and evaluated the PAE of 25 edible oil samples from supermarkets in Hangzhou city., Methods: The internal standard solution (D4-DEHP) was added in edible vegetable oil sample. The analytes were extracted by acetonitrile with 1 min vortex, and centrifuged at 3050×g for 5 min. The supernatant was then cleaned with silica/PSA column, and eluted with acetonitrile. The elution was dried with N2 flow at 50°C and diluted to 1.0 ml with hexane. Then, 17 PAE were tested by GC-MS and quantified with internal standards. The repeatability and sensitivity of the assay were evaluated. PAE were then determined in 25 plastic buckets of edible vegetable oil from supermarkets in Hangzhou city., Results: By the quantification of internal standard of D4-DEHP, a good linearity range of related 17 PAE was observed. The correlation coefficient was 0.994-1.000 and the standard lowest quantified level was 0.05-0.15 µg/ml. The spiking recoveries of 17 PAE were 78.3%-108.9% with the RSD of 4.3%-12.1% (n=6). The method detection limits were 0.1-0.2 mg/kg. In 25 plastic buckets of edible vegetable oil from Hangzhou, DMP, DEP, DIBP, DBP and DEHP were detected at the range of <0.1-1.8 mg/kg and the detection rates were 12% (3/25), 24% (6/25), 100% (25/25), 96% (24/25) and 100% (25/25), respectively. Other 12 PAE was not detected. For DBP with the level of <0.1 to 1.3 mg/kg, the results of 16% (4/25) samples exceeded the regular migrating limit of 0.3 mg/kg. For DEHP of <0.2-1.8 mg/kg, the data of 12% (3/25) samples were beyond the regular migrating limit of 1.5 mg/kg., Conclusion: The pretreatment by silica/PSA mixed solid phase extraction column can satisfy the PAE determination requirements in edible vegetable oils. The DMP, DIBP, DEP, DBP and DEHP were detected from the survey of 25 edible oil samples in Hangzhou city.

Objective: To observe the ethyl carbamate concentrations in different commercial fermented foods in Hangzhou in 2010., Methods: In 2010, 237 commercial fermented food samples of eight categories, including yellow wine, white spirit, wine, beer, cooking wine, sauce, vinegar and fermented bean curd, were purchased from 3 different size markets respectively in Hangzhou. The ethyl carbamate was measured by gas chromatography-mass spectrometry in selection ion mode, after the samples were coupled with D5-ethyl carbamate, and purified by diatomite solid phase extraction column., Results: The results showed that ethyl carbamate was detected in all samples analyzed (100%) with the range from 2.0 µg/kg to 515.0 µg/kg. The ethyl carbamate average (median) levels in 8 food categories were descending with fermented red bean curd (182.2 µg/kg (161.2 µg/kg)), yellow wine (159.6 µg/kg (121.0 µg/kg)), cooking wine (86.8 µg/kg (95.6 µg/kg)), white spirit (72.0 µg/kg (60.5 µg/kg)), soy sauce (47.2 µg/kg (40.7µg/kg)), vinegar (26.7 µg/kg (31.8 µg/kg)), wine (15.7 µg/kg (16.8 µg/kg)) and beer (2.2 µg/kg (2.3 µg/kg))., Conclusion: The ethyl carbamate was detected in all fermented foods in Hangzhou in 2010, and the levels of ethyl carbamate in red bean curd and yellow wine were higher than others.

Objective: To investigate clinical risky factors and improve the ability to discover critically ill patients suffering from influenza A H1N1 in emergency department., Methods: A retrospective study of 65 cases with severe influenza A H1N1 was conducted. Univariate analysis and multivariate Logistic regression analysis were used to identify independent predictors of critically ill cases. Goodness of Fit test was performed on an established clinical predicting scoring (prediction of critically ill with influenza score, PCIS) system., Results: Univariate analysis showed that significant difference between critically ill and severe group consisted of eight factors including age distribution, more than one comorbidity, chronic cardiovascular disease, oncologic diseases, sputum, myalgia/arthralgia, dyspnea and oxygen saturation at admission. A multivariate Logistic regression showed an association between development of critical illness and more than one comorbidity [odds ratio (OR)=7.611, 95% confidence interval (95%CI) 1.635-35.429], myalgia/arthralgia (OR=7.523, 95%CI 1.462-38.716), dyspnea (OR=11.090, 95%CI 1.373-89.565), and oxygen saturation<0.95 at admission (OR=8.088, 95%CI 1.019-84.969, all P<0.05). The prognostic criteria had a good discriminative ability [area under receiver operating characteristic curve (AUC) was 0.922, 95%CI 0.860-0.985, P=0.000]. The PCIS scoring system was established according to the level of four high risk factors. Low risk (PCIS 0-1), intermediate risk (PCIS 2), high risk (PCIS 3), and very high risk (PCIS≥4) were categorized for predicting the occurrence of critical illness, and the Goodness of Fit test was good (R(2)=0.940 6, P=0.030 1)., Conclusion: Emergency physician can predict the development of critical condition in patients with influenza A H1N1 by using clinical characteristics including comorbidity, myalgia/arthralgia, dyspnea, and oxygen saturation at admission, and it is helpful in making clinical decision.

This study is to explore whether the Wnt/beta-catenin signaling pathway is involved in the process of tripchlorolide (T4) protecting against oligomeric Abeta(1-42)-induced neuronal apoptosis. Primary cultured cortical neurons were used for the experiments on day 6 or 7. The oligomeric Abeta(1-42) (5 micromol x L(-1) for 24 h) was applied to induce neuronal apoptosis. Prior to treatment with Abeta(1-42) for 24 h, the cultured neurons were pre-incubated with T4 (2.5, 10, and 40 nmol x L(-1)), Wnt3a (Wnt signaling agonists) and Dkk1 (inhibitors) for indicated time. Then the cell viability, neuronal apoptosis, and protein levels of Wnt, glycogen synthase kinase 3beta (GSK3beta), beta-catenin and phospho-beta-catenin were measured by MTT assay, TUNEL staining and Western blotting, respectively. The result demonstrated that oligomeric Abeta(1-42) induced apoptotic neuronal cell death in a time- and dose-dependent manner. Pretreatment with T4 significantly increased the neuronal cell survival and attenuated neuronal apoptosis. Moreover, oligomeric Abeta(1-42)-induced phosphorylation of beta-catenin and GSK3beta was markedly inhibited by T4. Additionally, T4 stabilized cytoplasmic beta-catenin. These results indicate that tripchlorolide protects against the neurotoxicity of Abeta by regulating Wnt/beta-catenin signaling pathway. This may provide insight into the clinical application of tripchlorolide to Alzheimer's disease.

Aim: To explore the possible mechanism of tert-butyl hydroperoxide (t-BHP)-induced apoptosis in murine MIN6 pancreatic beta-cells., Methods: MIN6 cells were cultured in vitro. Cell damage was evaluated by epifluorescence microscopy after staining with AO-EB. The percentage of cell apoptosis was determined by flow cytometric assay after Annexin- V-PI staining. Nitric oxide levels were measured by Griess assay. Inducible nitric oxide synthase(iNOS) protein and NF-kappaBp65 fragment were detected by Western blot., Results: Exposure of 25 micromol/L t-BHP to MIN6 cells for 60 min, cell viability was reduced and the percentage of apoptosis was increased significantly. The levels of cytoplasmic iNOS protein and nitrite were elevated. Meanwhile, treatment with t-BHP resulted in nucleus NF-kappaBp65 fragment peaking at 20 min. Both L-NAME and N-Acetyl-l-cysteine (NAC) attenuated the elevated levels of nitrite and percentage of apoptosis due to t-BHP alone., Conclusion: NF-kappa-iNOS-nitric oxide signalling pathway can mediated t-BHP induced apoptosis in MIN6 cells .

Objective: To develop a simple, accurate, rapid, economic, large-scale detection method for the detection of single nucleotide polymorphisms (SNPs) metabolic enzymes, using polymerase chain reaction with confronting two-pair primers (PCR-CTPP)., Methods: The primers of CYP1A1 (A4889G), EPHX1 (A416G) and NQO1 (C609T) were designed for PCR-CTPP, and the PCR conditions were optimized. The results of genotyping were verified by DNA sequencing. The above SNPs were detected by the PCR-CTPP detection method in a randomly selected 183 healthy individuals of Han ethnicity. The genotype frequencies were analyzed and compared with people from other ethnicities., Results: The allele-specific bands of CYP1A1 (A4889G), EPHX1 (A416G) and NQO1 (C609T) were successfully amplified by PCR-CTPP under the optimal conditions and the results of genotyping were consistent with DNA sequencing. Among 183 healthy Han individuals, the genotypic distributions of CYP1A1 (A4889G) , EPHX1 (A416G) and NQO1 (C609T) showed that the wild-type, homozygous variants, and heterozygotes were 103 (56.3%), 8 (4.4%), 72 (39.3%) and 142 (77.6%), 4 (2.2%), 37(20.2%), 60(32.8%), 32 (17.5%), 91 (49.7%) respectively. The distributions of genotypes were all in accordance with the Hardy-Weinberg equilibrium (P > 0.05), with statistical differences and with other ethnic populations (P < 0.05)., Conclusion: The SNPs of metabolic enzymes can be detected by PCR-CTPP method which is simple, accurate, rapid, economic and with large scale. PCR-CTPP can be used for large scale clinical and epidemiological screening.

Extracts of Chinese herb Tripterygium wilfordii Hook. f. (TWHF) have been found to have potent anti-inflammatory and immunosuppressive functions and widely used in China for treatment of rheumatoid arthritis. Also they have been considered to be the potential drugs in the treatment of tumor and acute graft rejections. With the progress of neuroimmunological research on neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson disease (PD) and multiple sclerosis (MS), the neuroprotective strategies to rescue neurons from immunological injury are currently being explored. Recently, studies have indicated that extracts of TWHF such as triptolide, tripchlorolide and (5R)-5-hydroxytriptolide are able to attenuate progression of these neuroimmunologic disorders in vitro and in vivo. Accumulating evidence has shown that they can promote neuronal survival and neurite growth and facilitate functional recovery of brain injury by invoking distinct mechanisms that are related to their neuroprotective roles as inhibitor of neuroinflammatory toxicity of activated-microglia, antioxidants, calcium channel blockers, neurotrophic actions, modulating T cell functions, inhibitor of transcriptional activation of NF-kappaB on genes and signaling. Significant pharmaceutical strategies against neuroimmunologic disorders will hopefully be discovered by understanding the valuable components of TWHF.

To explore the effect of glucagon-like peptide-1 receptor agonist--Exendin-4 (Ex-4) on murine MIN6 pancreatic beta-cells apoptosis induced by oxidative stress, the morphological changes of cell damage were evaluated by epifluorescence microscopy after staining with AO-EB. The percentage of cell apoptosis was determined by flow cytometric assay after Annexin-V-FITC-PI staining. Nitric oxide level was measured by Griess reagent assay. Inducible nitric oxide synthase (iNOS) protein and NF-kappaBp65 fragment were detected by Western blotting. Ex-4 inhibited the increase of nitrite level and percentage of apoptosis induced by t-BHP in MIN6 cells. Furthermore, Ex-4 partly reduced the expression of iNOS protein and the ratio of NF-kappaBp65 protein in nucleus:cytosol induced by t-BHP. These results suggest that Ex4 protects MIN6 pancreatic kappa-cells from oxidative stress-induced apoptosis via down-regulation of NF-kappaB-iNOS-nitric oxide pathway.

This study is to explore the effect of ginsenoside Rb1 on the process of beta-amyloid peptide(25-35) (Abeta(25-35)) -induced hyperphosphorylation of tau protein, and on the level of cyclin-dependent kinase 5 activator, p25/p35. Western blotting and/or immunocytochemical staining were used to detect the levels of phosphorylation of tau protein at the sites of Thr205, Ser396, Ser404 in hippocampal neurons, cdk5 and p25/p35. After exposure to Abeta(25-35) (20 micromol x L(-1)) for 12 h, the levels of tau protein phosphorylation at the sites of Thr205, Ser396, Ser404 were enhanced, the level of p25 was increased, but the level of protein cdk5 was not changed markedly. Pretreatment with ginsenoside Rb1 reduced Abeta(25-35) -induced hyperphosphorylation of tau protein and decreased the lever of p25, but had no effect on cdk5. Ginsenoside Rb1 can attenuate Abeta(25-35) -induced hyperphosphorylation of tau protein through CDK5 signal pathway.

Objective: To identify the mutation of low density lipoprotein receptor(LDLR) gene in a large Chinese family with familial hypercholesterolemia(F H) and make a discussion on the pathogenesis of FH at the molecular level., Methods: Investigations were made on a patient with the clinical phenotype of homozygous FH and his parents for mutations of promoter and all 18 exons of LDLR gene. Screening was carried out using Touch down PCR and a g arose gel electrophoresis, combined with DNA sequence analysis. The results were compared with the normal sequences in GenBank and FH database (www.ucl.uk/fh) t o find the mutation. Then the mutation was identified in other members of the family. In addition, the authors screened the apolipoprotein B(100) (apoB(100)) gene f or known mutations (R3500Q) that cause familial defective apoB(100) (FDB) by PCR-RFLP., Results: A novel homozygous IN III 5' GT --> AT mutation in the splice donor of LDLR intron 3 was detected in the homozygote propositus with FH. The mutation was also identified in four heterozygous carriers in his family. No mutations R3500Q of apoB(100)were observed., Conclusion: A homozygous G --> A splice mutation in LDLR gene was first reported. The change of the splice donor in LDLR intron 3 may cause skipping of exon 3, which is responsible for FH. Perhaps it is a particular pathogenesis for Chinese people.