A Novel Multi-compartment Rotating Bioreactor for Improving ADSC--Spheroid Formation and its Application in Neurogenic Erectile Dysfunction.

OBJECTIVE: The aim of this study was to construct a multicompartment synchronous rotating bioreactor (MCSRB) for batch-production of homogenized adipose-derived stem cell (ADSC) microspheres and treat neurogenic erectile dysfunction (ED). METHODS: Firstly, an MCSRB was constructed using a centrifugal device and hinged trays. Secondly, influence factors (density, rotational speed) on the formation of ADSC-spheroids were explored. Finally, a neurogenic ED model was established to verify the effectiveness and safety of ADSC-spheroids for ED treatment. RESULTS: An MCSRB promoted ADSCs to gather microspheres, most of which were 90-130 µm in diameter. Supernatant from three-dimensional culture led to a significant increase in cytokine expression in ADSCs and migration rate in human umbilical vein endothelial cells (HUVECs) compared to control groups. The erectile function and pathological changes of the penis were improved in the ADSC-spheroids treatment group compared to the traditional ADSCs treatment group (p < 0.01). CONCLUSION: Efficient, batch, controlled and homogenized production of ADSC stem cell microspheres, and effective improvement of erectile dysfunction in neurogenic rats can be achieved using the MCSRB device.

Role of mechanotransduction mediated by YAP/TAZ in the treatment of neurogenic erectile dysfunction with low-intensity pulsed ultrasound.

BACKGROUND: Erectile dysfunction (ED) and weakness of the penis are processes related to hemodynamic alteration. Low-intensity pulsed ultrasound (LIPUS), as a new mechanical modality for the treatment of ED, deserves to be explored in depth for the biomechanical mechanisms it exerts. OBJECTIVE: The aim of this study was to explore the role of YAP/TAZ-mediated mechanotransduction in mechanical therapy for the treatment of neurogenic erectile dysfunction (NED). MATERIALS AND METHODS: Forty-two male SD rats (12 w old) were randomly divided into sham-operated (n = 14), bilateral cavernous nerve injury (BCNI, n = 14), and LIPUS-treated (n = 14) groups. Intracavernosal pressure/mean arterial pressure (ICP/MAP) was measured 14 and 28 days after treatment. Penile tissue specimens were collected for pathological examination, and the changes in YAP, TAZ, connective tissue growth factor (CTGF), CYR61, LATS1, and p38 mitogen-activated protein kinase expression levels were assessed by Western blot, real-time quantitative polymerase chain reaction (RT-qPCR) and immunological staining. RESULTS: Compared with BCNI, LIPUS significantly improved ICP/MAP levels and enhanced histopathological changes. The penile expression levels of YAP, TAZ, CTGF, and CYR61 were significantly downregulated in the BCNI group (p < 0.01), and LIPUS upregulated the expression levels of these proteins (p < 0.05). The expression levels of p-LATS1 and LATS1 were not significantly different among the groups (p > 0.05). Interestingly, the expression level of p-p38/p38 significantly increased in BCNI rats (p < 0.05), which was reversed by LIPUS treatment (p < 0.05). However, the p38 inhibitor SB203580 did not change the expression of YAP/TAZ in rat primary smooth muscle cells or mouse MOVAS cells (p > 0.05). DISCUSSION AND CONCLUSION: LIPUS can effectively improve penile erectile function in NED rats. The underlying mechanism may be related to the regulation of YAP/TAZ-mediated mechanotransduction. However, the upstream regulatory signal may differ from the classical Hippo pathway.

Dissolved biochar fractions and solid biochar particles inhibit soil acidification induced by nitrification through different mechanisms.

Biochar can inhibit soil acidification by decreasing the H+ input from nitrification and improving soil pH buffering capacity (pHBC). However, biochar is a complex material and the roles of its different components in inhibiting soil acidification induced by nitrification remain unclear. To address this knowledge gap, dissolved biochar fractions (DBC) and solid biochar particles (SBC) were separated and mixed thoroughly with an amended Ultisol. Following a urea addition, the soils were subjected to an incubation study. The results showed that both the DBC and SBC inhibited soil acidification by nitrification. The DBC inhibited soil acidification by decreasing the H+ input from nitrification, while SBC enhanced the soil pHBC. The DBC from peanut straw biochar (PBC) and rice straw biochar (RBC) decreased the H+ release by 16 % and 18 % at the end of incubation. The decrease in H+ release was attributed to the inhibition of soil nitrification and net mineralization caused by the toxicity of the phenols in DBC to soil bacteria. The abundance of ammonia-oxidizing bacteria (AOB) and total bacteria decreased by >60 % in the treatments with DBC. The opposite effects were observed in the treatments with SBC. Soil pHBC increased by 7 % and 19 % after the application of solid RBC and PBC particles, respectively. The abundance of carboxyl on the surface of SBC was mainly responsible for the increase in soil pHBC. Generally, the mixed application of DBC and SBC was more effective at inhibiting soil acidification than their individual applications. The negative impacts of dissolved biochar components on soil microorganisms need to be closely monitored.

[Electroencephalogram Features of Anti-N-Methyl-D-Aspartate Receptor Encephalitis and Their Value for Clinical Assessment].

Objective: To analyze the electroencephalogram (EEG) features of anti-N-methyl-D-aspartate receptor encephalitis (anti-NMDARE) and to study the clinical assessment value of the degree of EEG background slowing and the presence of δ brush. Methods: We enrolled 52 patients with anti-NMDARE and collected their clinical data, including age, sex, form of disease onset, status of tumor comorbidity, auxiliary examination findings (cerebrospinal fluid [CSF] anti-methyl-D-aspartate receptor antibody titers, magnetic resonance imaging [MRI] reports, and EEG results), treatment status, and follow-up after discharge. The degree of EEG background abnormality and the presence of δ brush in the EEG of patients with different clinical features were analyzed. Results: Among the 52 patients, 7 (14%) had normal EEG, and 45 (87%), abnormal EEG, including 25 (48%) with mild abnormalities, 11 (21%) with moderate abnormalities, and 9 (17%) with severe abnormalities. δ brush was seen in 6 (12%) patients. At the time of EEG, 32 (62%) patients were in the mild condition group and 20 (38%) patients were in the severe condition group. After 1 year of follow-up, there were 45 (86%) patients in the good prognosis group and 7 (14%) patients in the poor prognosis group. The exacerbation of EEG background abnormalities and the presence of δ brush were indications for an increase in the proportion of patients who were in severe condition, who needed ICU admission, and who had poor prognosis ( P<0.01). The worse the EEG background abnormalities, the higher the proportion of CSF antibody titers>1∶10 ( P=0.035), and the higher the proportion of patients initiating second-line immunotherapy ( P=0.008). The δ brush was seen a higher proportion in patients with comorbid tumors ( P=0.012). The probability of δ brush presence was higher in the first-time diagnosis cases than that in recurrent cases ( P=0.023). Conclusions: The degree of EEG slowing and the presence of δ brush have shown consistent performance in assessing patients' condition and predicting prognosis. The slower the EEG, the more severe the disease, and the worse the prognosis. The presence of δ brush indicates severe disease and poor prognosis. EEG slowing is correlated with the immune status of patients with anti-NMDARE. The slower the EEG, the more severe the immune abnormalities. In clinical practice, patient EEG should be under dynamic monitoring in order to evaluate the effect of immunotherapy. If EEG slowing is not improved, enhanced immunotherapy should be considered as early as possible. The δ brush is seen at a higher proportion in patients with comorbid tumors. Therefore, active efforts should be made to screen for tumors when δ brush is present.

Application of measuring electrochemical characteristics on plant root surfaces in screening Al-tolerant wheat.

To explore the relationship between Al phytotoxicity and the electrochemical characteristics of wheat root surfaces, a new chemical mechanism for tolerance of wheat to Al toxicity was initially proposed by conducting acute root elongation experiment, adsorption/desorption experiment, streaming potential determination, and infrared spectroscopy (ATR-FTIR) analysis respectively to classify the grade of Al tolerance of 92 wheat cultivars and quantitatively characterize the electrochemical properties of their root surfaces. Then a pot experiment was conducted with the screened wheat cultivars with different Al resistance grown on acid soils to verify their tolerance to Al toxicity. Results show that zeta potentials of the roots of 67 wheat cultivars at pH4.46 were significantly negatively correlated with Al(Ⅲ) adsorbed on the roots and their relative root elongation (P < 0.05), indicating that wheat roots with less negative charges is more tolerant to Al toxicity. Based on the mechanism, 14 Al-tolerant, 23 medium Al-tolerant and 30 Al-sensitive wheat cultivars were classified. The pot experiment reveals that the relative dry weight of Al-tolerant wheat cultivars was generally greater than that of medium Al-tolerant and Al-sensitive wheat cultivars and Al-tolerant wheat cultivars accumulate less Al in their shoots, which further verifies the relationship among charge characteristics, tolerance of wheat to Al toxicity, and Al uptake by wheat. The negative charges derived from organic functional groups on root surfaces could influence the exchangeable and complexed Al(Ⅲ) adsorbed on wheat roots and thereby affect Al tolerance of wheat cultivars. This finding not only provides a new perspective to screen Al-tolerant wheat cultivars and explain the mechanism of tolerance of wheat to Al toxicity, but is also useful for the prediction of differences in the uptake of Al in the shoots between Al-tolerant and Al-sensitive wheat cultivars, and finally contributes to the prevention of food security risk caused by Al in acid soils.

Effects of straw decayed products of four crops on the amelioration of soil acidity and maize growth in two acidic Ultisols.

Variable charge soils have low agricultural productivity associated with low pH, low cation exchange capacity (CEC), and low pH buffering capacity (pHBC). As a result of rapid acidification rates, these soils are prone to infertility resulting from Al phytotoxicity and deficiency of P, Ca, Mg, and K, and thus require amendments that can ameliorate soil acidity and enhance soil CEC and pHBC. A 30-day pot experiment was carried out using a clay Ultisol and a sandy Ultisol amended with straw decayed products (SDPs) of peanut, pea, canola, and rice. The results showed that applying SDPs increased the soil CEC, organic matter content, and exchangeable base cations in the two Ultisols. The ameliorative effects of the SDPs were superior for the sandy Ultisol than for the clay Ultisol. The addition of SDPs significantly increased soil pH and pHBC of the two Ultisols, and simultaneously decreased soil exchangeable Al3+. Among them, the greatest effect was found in the treatment with pea straw decayed products (PeaSD). The soil pHs of clay Ultisol and sandy Ultisol treated with PeaSD were respectively 5.70 and 7.37 and were 1.26 and 2.63 pH units higher than those of control. Also, applying SDPs increased maize seedling biomass in both soils and the most significant effect was found in the treatment with PeaSD, which were 0.97 (clay Ultisol) and 2.5 (sandy Ultisol) times higher than in the respective controls. The results of this study demonstrated that carefully selected straws for SDP production can effectively improve soil chemical properties, enhanced soil pHBC, and thus promote agricultural sustainability.

Phytotoxicity of Cu2+ and Cd2+ to the roots of four different wheat cultivars as related to charge properties and chemical forms of the metals on whole plant roots.

The relationship between the chemical forms of Cu2+ and Cd2+ adsorbed on the roots of different wheat cultivars and their phytotoxic effects on the plants were investigated. The wheat varieties Dunmaiwang (DMW), Tekang 6 (TK6), Zhongmai895 (ZM895), and Chaojixiaomai (AK68) were used. The zeta potentials of wheat roots, measured by the streaming potential method, were used to characterize root charge properties. Results indicated that the changes in zeta potential at pH 4.01-6.61 were 14.7, 15.53, 13.01, and 12.06 mV for ZM895, AK68, DMW, and TK6, respectively. The negative charge and functional groups on ZM895 and AK68 roots were greater than on DMW and TK6 roots, which led to more exchangeable and complexed Cu2+ and Cd2+ on ZM895 and AK68 roots and increased Cu2+ and Cd2+ toxicity compared to DMW and TK6. Coexisting cations, such as Ca2+, Mg2+, K+, and NH4+, alleviated Cu2+ and Cd2+ toxicity to wheat roots through competition for adsorption sites on the roots, which decreased exchangeable and complexed Cu2+ and Cd2+ on wheat roots. The Ca2+ and Mg2+ were most effective in alleviating heavy metal toxicity and they decreased exchangeable Cu2+ on AK68 roots by 39.14% and 47.82%, and exchangeable Cd2+ by 8.51% and 28.23%, respectively.

Biochar retards Al toxicity to maize (Zea mays L.) during soil acidification: The effects and mechanisms.

Biochar can effectively alleviate the Al phytotoxicity in acidic soils due to its alkaline nature. However, the longevity of this alleviation effect of biochar under re-acidification conditions is still unclear. In the present study, the maize root growth responding to the simulated re-acidification of two acidic soils amended by peanut straw biochar or Ca(OH)2 was investigated to evaluate the long-term effect of biochar on alleviating Al toxicity in acidic soils. Compared with Ca(OH)2 amendment, the application of biochar significantly retarded Al toxicity to plant during soil re-acidification. When 4.0 mM HNO3 was added, the maize seedling root elongation in an Oxisol with biochar was 99% higher than that in the Oxisol with Ca(OH)2. Also, the Evans blue uptake and Al content in the root tip in the biochar treatment were 60% and 51% lower than those in the Ca(OH)2 treatment. The retarding effect was mainly attributed to the slow decrease in soil pH during acidification and the release of dissolved organic carbon (DOC) in the soils amended by biochar. The slower decrease in soil pH resulting from the increased pH buffering capacity after biochar application inhibited the increase of soluble and exchangeable Al during re-acidification. The increased DOC after biochar application decreased the toxic soluble Al speciation at the same pH value and total Al concentration in soil solution. Therefore, given the re-acidification of soils, biochar presented a longer-term effect on alleviating Al toxicity of acidic soil than liming.

Effects of extracellular polymeric substances of Pseudomonas fluorescens, citrate, and oxalate on Pb sorption by an acidic Ultisol.

The continuous production of low molecular weight (LMW) organic acids by plants and microorganisms coupled with the continuous presence of extracellular polymeric substances (EPS) in soils is a guarantee that the mobility of heavy metals in soils will be controlled. The effects of citrate, oxalate, and EPS on the adsorption of Pb by an acidic Ultisol were studied both as a function of pH and ionic strength. Electrokinetic potential measurements were also employed to observe to what extent each ligand affected the surface charge property of the Ultisol. All the ligands shifted the zeta potential of the Ultisol to the negative direction, implying that the surface charge of the soil became more negative. The effect on the zeta potential of the soil was observed in the order of oxalate ˃ citrate ˃ EPS. The quantity of Pb adsorbed at each pH (3.0-7.0) reflected the corresponding change in the zeta potential as induced by each ligand. The presence of the ligands shifted the isoelectric point of the Ultisol from 4.8 to 3.2 for the EPS system and below 3.0 for the citrate and oxalate systems. More Pb was adsorbed in the presence of oxalate than in the presence of citrate and EPS. The two most outstanding mechanisms that governed the adsorption of Pb by the Ultisol were (1) electrostatic attraction which was supported by the increase in negative zeta potential of the Ultisol and, (2) complexation which was supported by the lesser proportion of Pb adsorbed in the citrate system at higher pH and also by the spectroscopic data for EPS. The combination EPS + citrate + oxalate was more effective in enhancing the adsorption of Pb than the combination EPS + oxalate and EPS + citrate.

[Optimized procedure for fluorescence in situ hybridization in rapid prenatal diagnosis of common aneuploidy].

OBJECTIVE: To optimize the procedure of fluorescence in situ hybridization (FISH), and evaluate it in rapid prenatal diagnosis of common aneuploidy. METHODS: Amniotic fluid samples from 300 pregnant women were tested by both interphase FISH and conventional cell culture for karyotyping from September 2009 and September 2010. RESULTS: Seven cases of trisomy 21, 4 of trisomy 18, 2 of monosomy X, 1 of XXY, 1 of XXX, and 1 of triploidy were detected by FISH in the 300 amniotic fluid samples. It was concordant with the results from conventional karyotype analysis. The concordance rate was 100%. CONCLUSION: Through a technical modification of FISH procedure, the detection accuracy and specificity was not affected but testing cost reduced greatly. It can be used in rapid prenatal diagnosis of common aneuploidy.

[Evaluation on parallel HER-2 testing using in situ hybridization and immunohistochemistry in breast cancer tissues].

OBJECTIVE: To evaluate the application of the immunohistochemistry (IHC) and the fluorescence in situ hybridization (FISH) in detecting the amplification and the expression of HER-2 gene in the breast cancer patients. METHODS: Sixty-six cases of paraffin-embeded breast cancer samples with overexpression, low or no expression of HER-2 gene as detected by IHC were analyzed for HER-2 gene amplification using FISH. RESULTS: Among the 42 samples with HER-2 gene overexpression (3+/2+) detected by IHC, 31 showed positive HER-2 gene amplification and 11 showed negative HER-2 gene amplification in FISH. In the 24 samples with low or no HER-2 gene expression (1+/-) detected by IHC, no HER-2 gene amplification was detected by FISH. The results of the two testing methods showed a good consistency with the kappa coefficient of 0.672 (P<0.001). We also found that the 17 chromosome polysomy in 42% of the samples and the incidence of 17 polysomy was significantly higher in the HER-2 gene overexpression (3+/2+) group than in low or no HER-2 gene expression (1+/-) group (chi(2)=4.688, P=0.03). CONCLUSION: IHC can be used as a screening method for detecting HER-2 gene amplification, and FISH should be performed in cases of HER-2 gene overexpression (3+/2+) as detected by IHC.

[Second trimester maternal serum screening for Down's syndrome in women of advanced maternal age: a multi-center prospective study].

OBJECTIVE: To evaluate the performance characteristics of the second trimester double test for the detection of fetal Down's syndrome (DS) in women of advanced maternal age (AMA). METHODS: We undertook a prospective nation-wide multi-centered study and chose alpha-fetoprotein (AFP) and free beta-subunit of human chorionic gonadotrophin (free beta-hCG) as the serum markers. Between May 2004 and September 2006, 12 centers participated in the collection and analysis of maternal serum AFP and free beta-hCG. Patients with an increased risk of DS (> or = 1/270) were offered genetic amniocentesis. Follow up of the outcome of all pregnancies was obtained. Patients were divided into two groups, the AMA group and the non-AMA group and the screening efficiency was evaluated in both groups. RESULTS: A total of 66 132 singleton pregnancies were included in the study, and there were 3610 (5.46%) AMA women. The median maternal age of AMA women was 36.8years (35 - 47 years). At a cut-off of 1/270, in the AMA group, the number of positive cases screened was 727 and 22 cases of fetal DS were detected; the number of negative cases screened was 2883, and no fetal DS was found. In the non-AMA group, the number of positive cases screened was 4743 and 69 cases of fetal DS were detected; the number of negative cases screened was 57 779, and 6 cases of fetal DS were diagnosed postnatally. In AMA group, the detection rate (DR), false positive rate (FPR) and odds of being affected given a positive result (OAPR) were 100%, 19.7% and 3.0% respectively. In the non-AMA group, the DR, FPR and OAPR were 92.0%, 7.5% and 1.5% respectively. CONCLUSION: The double-marker test using AFP and free beta-hCG is an effective screen strategy for second-trimester detection of Down syndrome in AMA women.

[Second trimester maternal serum screening for Down's syndrome in mainland China: a multi-center prospective study].

OBJECTIVE: To evaluate the performance characteristics of the second trimester double-marker test for the detection of fetal Down's syndrome in mainland China. METHODS: This prospective national multi-centered study used alpha-fetoprotein (AFP) and free beta-subunit of human chorionic gonadotrophin (free beta-hCG) as the serum markers. From May 2004 to September 2006, 11 centers participated in the collection and analysis of maternal serum AFP and free beta-hCG between 14 and 20(+6) weeks of pregnancy. The screening results were calculated using the standard algorithm based on the standard database provided with the analytic software. Patients with an increased risk of Down's syndrome pregnancy (> or = 1/270) were offered genetic amniocentesis. Outcomes of all pregnancies were obtained. RESULTS: A total of 66 132 singleton pregnancies were included in the study. The median maternal age was 27 years. At a cut-off of 1 in 270, the detection rate (DR) based on a Caucasian database was 72% corresponding to a false positive rate (FPR) of 5%, and the DR based on the Chinese database was raised to 76% corresponding to an FPR of 5%. CONCLUSION: The double-marker test using AFP and free beta-hCG is an effective screen strategy for second-trimester detection of fetal Down's syndrome in mainland China. Ethnic variance exists between the Caucasian and Chinese populations. The accuracy of screening is increased by the use of race-specific medians.