Evaluation of sperm mitochondrial function using rh123/PI dual fluorescent staining in asthenospermia and oligoasthenozoospermia

Objective：The recent advent of flow cytometry（FCM）,coupled with fluorescent dyes,has been successfully applied to assess mitochondrial function.The aim of this study was to investigate the feasibility and clinical significance of detecting sperm mitochondrial function and to evaluate sperm mitochondrial function by using Rhodamine 123/propidium（Rh123/PI）dual fluorescent staining and FCM in asthenospermia and oligoasthenozoospermia.Methods：Twenty-five fertile men（with normal sperm parameters）and 230 infertile patients were examined.Fifty-five patients of the above 230 patients were selected for idiopathic infertility samples and were divided into two groups：asthenospermia（n=30）and oligoasthenozoospermia（n=25）.Rh123/PI dual fluorescent staining and FCM were carried out to examine sperm mitochondrial function.Results：Significant differences were found between the normal and abnormal semen samples（P0.05）when Rh123＋/PI-,Rh123-/PI＋and Rh123-/PI-sperm were examined by FCM,but there was no significant difference between the asthenospermia（P=0.469） and oligoasthenozoospermia group（P=0.950）when Rh123＋/PI-and Rh123-/PI＋sperm were then examined;however,a significant difference was found between the 2 groups（P=0.003）when Rh123-/PI-sperm were examined.There was no correlation between Rh123-/PI-sperm and semen parameters in the normal group,but there was a significant negative correlation between the sperm concentration and Rh123-/PI-sperm in asthenospermia and oligoasthenozoospermia patients（r=-0.509,-0.660;P=0.018,0.038）.Conclusion：Rh123/PI dual fluorescent staining and FCM can provide reliable information to assess the quality of sperm and reveal differences in mitochondrial membrane potential in asthenospermia and oligoasthenozoospermia.

Identifying the major fluorescent components responsible for ultrafiltration membrane fouling in different water sources

Three-dimensional fluorescence excitation–emission matrix（EEM） coupled with parallel factor analysis（PARAFAC） was performed for a total of 18 water samples taken from three water sources（two lakes and one wastewater treatment plant（WWTP） secondary effluent）,with the purpose of identifying the major ultrafiltration（UF） membrane foulants in different water sources. Three fluorescent components（C1, C2 and C3） were identified,which represented terrestrially derived humic-like substances（C1）, microbially derived humic-like substances（C2）, and protein-like substances（C3）. The correlations between the different fluorescent components and UF membrane fouling were analyzed. It was shown that for the WWTP secondary effluent, all three components（C1, C2 and C3） made a considerable contribution to the irreversible and total fouling of the UF membrane.However, for the two lakes, only the C3 exhibited a strong correlation with membrane fouling, indicating that the protein-like substances were the major membrane foulants in the lake waters. Significant attachment of C1, C2 and C3 to the UF membrane was also confirmed by mass balance analyses for the WWTP secondary effluent; while the attachment of C1 and C2 was shown to be negligible for the two lakes. The results may provide basic formation for developing suitable fouling control strategies for sustainable UF processes.

A Fiber Optic Sensor for 2-cholrophenol Analysis based on Oxygen Sensing System

A fiber optic 2-cholrophenol（2-CP） sensor was developed based on the fluorescence quenching of molecular oxygen on the oxygen-sensitive membrane and O2 consumption during catalytic oxidation reaction of 2-CP. The 2-CP concentration can be determined by utilizing a lock-in amplifier to measure the change in the fluorescence lifetime of an oxygen-sensitive membrane, in which the tris（2,2′-bipyridyl） ruthenium（II） chloride complexes（Ru（II）（byp）3Cl2） were immobilized in cellulose acetate（CA） via simple hybridized approach. The experimental results show the good linear relationship between the phase delay of sensitive membrane and 2-CP concentration in its detection range of 1×10-7 to 1×10-5 mol/L and 1×10-5 to 1×10-4 mol/L. The detection limit of the sensor is 7×10-8 mol/L（S/N=3） and the response time is 5 min. Our experimental measurements confirmed good response characteristics of the as-prepared fiber optic 2-CP sensor, as well as its capability to detect the 2-CP concentration in practical water samples.

Advanced treatment of municipal wastewater by nanofiltration： Operational optimization and membrane fouling analysis

Municipal sewage from an oxidation ditch was treated for reuse by nanofiltration（NF） in this study. The NF performance was optimized, and its fouling characteristics after different operational durations（i.e., 48 and 169 hr） were analyzed to investigate the applicability of nanofiltration for water reuse. The optimum performance was achieved when transmembrane pressure = 12 bar, p H = 4 and flow rate = 8 L/min using a GE membrane. The permeate water quality could satisfy the requirements of water reclamation for different uses and local standards for water reuse in Beijing. Flux decline in the fouling experiments could be divided into a rapid flux decline and a quasi-steady state. The boundary flux theory was used to predict the evolution of permeate flux. The expected operational duration based on the 169-hr experiment was 392.6 hr which is 175% longer than that of the 48-hr one. High molecular weight（MW） protein-like substances were suggested to be the dominant foulants after an extended period based on the MW distribution and the fluorescence characteristics. The analyses of infrared spectra and extracellular polymeric substances revealed that the roles of both humic- and polysaccharide-like substances were diminished, while that of protein-like substances were strengthened in the contribution of membrane fouling with time prolonged. Inorganic salts were found to have marginally influence on membrane fouling. Additionally, alkali washing was more efficient at removing organic foulants in the long term, and a combination of water flushing and alkali washing was appropriate for NF fouling control in municipal sewage treatment.