Study on the mechanism of Fufang E'jiao Jiang on precancerous lesions of gastric cancer based on network pharmacology and metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufang E'jiao Jiang (FEJ) is a prominent traditional Chinese medicine prescription, which consists of Asini Corii Colla (Donkey-hide gelatin prepared by stewing and concentrating from the hide of Equus asinus Linnaeus., ACC), Codonopsis Radix (the dried roots of Codonopsis pilosula (Franch.) Nannf., CR), Ginseng Radix et Rhizoma Rubra (the steamed and dried root of Panax ginseng C.A. Mey., GRR), Crataegi Fructus (the mature fruits of Crataegus pinnatifida Bunge., CF), and Rehmanniae Radix Praeparata (the steamed and sun dried tuber of Rehmannia glutinosa (Gaertn.) Libosch. ex Fisch. & C.A. Mey., RRP). It is a popularly used prescription for "nourishing Qi and nourishing blood". AIM OF THE STUDY: To explore the potential mechanism of FEJ on precancerous lesion of gastric cancer in rats by combining network pharmacology and metabolomics. METHODS: Traditional Chinese Medicine Systems Pharmacology and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine were used to identify the ingredients and potential targets of FEJ. GeneCards database was used to define PLGC-associated targets. We built a herb-component-disease-target network and analyzed the protein-protein interaction network. Underlying mechanisms were identified using Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. In addition, 40% ethanol, N-methyl-N'-nitro-N-nitroguanidine and irregular eating were used to establish PLGC rats model. We also evaluated the efficacy of FEJ on MNNG-induced PLGC rats by body weight, histopathology, blood routine and cytokine levels, while the predicted pathway was determined by the Western blot. Ultra-performance liquid chromatography-tandem mass spectrometry-based serum non-targeted metabolomics was used to select potential biomarkers and relevant pathways for FEJ in the treatment of PLGC. RESULTS: Network pharmacology showed that FEJ exhibited anti-PLGC effects through regulating ALB, TNF, VEGFA, TP53, AKT1 and other targets, and the potential pathways mainly involved cancer-related, TNF, PI3K-AKT, HIF-1, and other signaling pathways. Animal experiments illustrated that FEJ could suppress inflammation, regulate gastrointestinal hormones, and inhibit the expression of PI3K/AKT/HIF-1α pathway-related proteins. Based on serum non-targeted metabolomics analysis, 12 differential metabolites responding to FEJ treatment were identified, and metabolic pathway analysis showed that the role of FEJ was concentrated in 6 metabolic pathways. CONCLUSION: Based on network pharmacology, animal experiments and metabolomics, we found that FEJ might ameliorate gastric mucosal injury in PLGC rats by regulating gastrointestinal hormones and inhibiting inflammation, and its mechanism of action is related to the inhibition of excessive activation of PI3K/AKT/HIF-1α signaling pathway and regulation of disorders of body energy metabolism. This comprehensive strategy also provided a reasonable way for unveiling the pharmacodynamic mechanisms of multi-components, multi-targets, and multi-pathways in Traditional Chinese Medicine.

[Nitrogen and phosphorus dynamics along the primary succession in the Hailuogou Glacier retreat area, eastern Tibetan Plateau, China.] / 青藏高原东缘海螺沟冰川退缩区植被原生演替的氮磷动态.

In this study, seven sampling sites (glaciers retreated for 0, 10, 30, 40, 50, 80 and 127 years) were chosen along a primary succession sequence in the Hailuogou glacial retreat area in the eastern margin of the Tibetan Plateau, China. The accumulation and cycling characteristics of N and P under different succession stages were analyzed by measuring biomass and N and P contents in surface soil and each vegetation layer. The N and P contents in leaves, branches and roots of tree layers decreased along the succession sequence, whereas the N and P contents in stems were higher in the late succession stage. The changes of N and P contents in litter and soil O layer were consis-tent with those in the leaves and branches of tree layers. Ecosystem N and P storage increased along the succession sequence. Ecosystem N accumulation was mainly dependent on the vegetation layer in the early succession stage. After the community reached the climax, soil became the main N pool of the ecosystem. Vegetation P storage was higher than that in the surface soil after 80 years of glacial retreat. The nutrient accumulation rate in each layer of the ecosystem was rapid in the middle succession stage, with an order of surface soil > tree layer > understory vegetation layer. The nutrient cycling coefficients of N and P in broadleaved forest in the middle stage were higher than those in coniferous forest in the late stage, whereas the N and P utilization efficiency was lower than that in coniferous forest. Therefore, the mechanism of low nutrient cycling and high utilization efficiency of coniferous trees was conducive to the their competition with other species, thus finally forming the climax community.

An Antisense Oligonucleotide Drug Targeting miR-21 Induces H1650 Apoptosis and Caspase Activation.

Non-small cell lung cancer is the most common malignant tumor in the world. Currently, chemotherapy is still the major method for non-small cell lung cancer treatment, but the problem of cancer drug resistance still exists, so we designed 5 different phosphorothioate oligonucleotides to silence key genes in tumor cell development, which could help avoid inducing cancer cell drug resistance. MicroRNAs have been shown to play a crucial role in the pathogenesis and progression of many malignancies, such as breast, colon, lung, and pancreatic cancer. According to the data from the Gene Expression Omnibus database, miR-21 has been reported to be one of the top 20 differentially expressed microRNAs screened using the Morpheus online tool, and miR-21 has been revealed to regulate a series of biological behaviors in cancer cells, including cell proliferation, migration, invasion, metastasis, and apoptosis. In recent years, gene therapy has emerged as a new therapeutic strategy for cancer treatment. Antisense oligonucleotides have recently been suggested as a novel approach for targeting microRNAs by antisense-based gene silencing. Five phosphorothioate oligonucleotides were designed, synthesized, and screened for anticancer activity. Reverse transcription-polymerase chain reaction was used to detect the relative expression of miR21. Among these 5 sequences, only phosphorothioate oligonucleotide 4 inhibited the proliferation of H1650 cells, and this effect was due to the induction of cancer cell apoptosis by activating the caspase-8 apoptotic pathway. In conclusion, this research confirmed the anticancer activity of phosphorothioate oligonucleotide 4 and revealed the underlying mechanism, which has the potential to be a novel anticancer strategy.

Asymmetric valley polarization and photoluminescence in MoS2/MoO3 heterostructure.

We investigate circularly polarized photoluminescence (PL) in the MoS2/MoO3 heterostructure, which was fabricated by transferring MoS2 monolayer to cover the MoO3 few layers on the SiO2/Si substrate. It is shown that the PL with the same helicity as the excitation light is dominant due to the inherent chiral optical selectivity, which allows exciting one of the valleys in MoS2 monolayer. The degree of polarization (DP), which characterizes the intensity difference of two chiral components of PL, is unequal for the right-handed and left-handed circularly polarized excitations in the MoS2/MoO3 heterostructure. This effect is different from the one in pristine MoS2. Our Raman spectra results together with ab initio calculations indicate the p-doped features of the MoS2 when it covers the MoO3 layers. Thus the possible explanation of the unequal DP is that the p-doping process generates a built-in voltage and therefore brings the difference of electron-hole overlaps between K and K' valleys. Namely the asymmetric valley polarization may be obtained in the MoS2/MoO3 heterostructure. Consequently, the circularly polarized PL caused by the electron-hole recombination at K and K' valleys manifests unequal DP for the right-handed and left-handed helix excitations. This asymmetric effect is further enhanced by decreasing the temperature in the MoS2/MoO3 heterostructure. Our investigation provides a unique platform for developing novel two-dimensional valleytronic devices.

Long-term sky islands generate highly divergent lineages of a narrowly distributed stream salamander (Pachyhynobius shangchengensis) in mid-latitude mountains of East Asia.

BACKGROUND: Climate oscillation may have a profound effect on species distributions, gene flow patterns and population demography. In response to environmental change, those species restricted to montane habitats experienced expansions and contractions along elevation gradients, which can drive differentiation among sky islands. RESULTS: The Shangcheng stout salamander (Pachyhynobius shangchengensis) is a cool stream amphibian restricted to high-elevation areas in the Dabie Mountains, East China. In the present study, we used mtDNA genes (Cyt b and ND2) of 193 individuals and 12 nuclear microsatellite loci genotyped on 370 individuals, representing 6 populations (JTX, KHJ, MW, TTZ, BYM and KJY) across the taxon's distribution area, to investigate their genetic variation and evolutionary history of P. shangchengensis. Most populations showed unusually high levels of genetic diversity. Phylogenetic analyses revealed five monophyletic clades with divergence times ranging from 3.96 to 1.4 Mya. Accordingly, significant genetic differentiation was present between these populations. Bayesian skyline plot analyses provided that all populations underwent long-term population expansions since the last inter-glacial (0.13 Mya ~ 0.12 Mya). Msvar analyses found recent signals of population decline for two northern populations (JTX and KHJ) reflecting a strong bottleneck (approximately 15-fold decrease) during the mid-Holocene (about 6000 years ago). Ecological niche modelling has shown a discontinuity in suitable habitats for P. shangchengensis under different historical climatic conditions. CONCLUSIONS: Our results suggest that the niche conservatism of P. shangchengensis and sky island effects may have led to long-term isolation between populations. In sky island refuges, the mid-latitude Dabie Mountains have provided a long-term stable environment for P. shangchengensis, which has led to the accumulation of genetic diversity and has promoted genetic divergence.

Photon-induced carrier recombination in the nonlayered-structured hybrid organic-inorganic perovskite nano-sheets.

The hybrid organic-inorganic perovskites (HOIPs) have attracted much attention recently due to their preeminent efficiency in solar cells. According to the difference on the crystalline structure, the HOIPs could be classified into layered and non-layered perovskites. Very recently, it has been realized that the non-layered HOIPs with common-vertex structure possess even better opto-electrical performance. Yet the carrier recombination mechanism in perovskite remains not very clear, and a clear understanding of this mechanism is essential to pinpoint the working mechanism of photovoltaic and electroluminescent materials. Here we report the optical studies on the hybrid perovskite crystalline nano-sheet of CH3NH3PbBr3 with common-vertex structure. It is shown that the non-layered perovskite crystalline nanosheets possess the exciton binding energy about two orders of magnitude smaller than that of the layered perovskite and the colloidal nanoplates, which is beneficial for the designing of the high-efficiency photovoltaic devices. By measuring the temperature-dependent photoluminescence (PL) spectra, the excitation-power-variant PL spectra, and the time-resolved PL spectra, we identify that both the free-carrier and the localized exciton recombination channels may coexist in the crystallites. Further, for the thin crystallite (∼60 nm), the free-carrier recombination channel dominates; whereas when the thickness increases beyond 200 nm, the localized exciton recombination channel plays the major role. We suggest these results are helpful to improve further the photovoltaic and electroluminescent performances of perovskite devices.

Hybrid coupling enhances photoluminescence of monolayer MoS2 on plasmonic nanostructures.

The efficiency of photoluminescence (PL) of transition-metal dichalcogenides (TMDCs) significantly influences their practical applications in optoelectronic devices. In this work, we study multiple coupling among excitons, surface plasmons, and optical modes, and their effects on PL of monolayer MoS2 atop plasmonic nanohole arrays. Under the illumination of visible light, strong intensity enhancement of PL from monolayer MoS2 is observed in the system. We further demonstrate that there exist excitons induced from MoS2, localized and propagating surface plasmons excited from nanoholes, and optical modes related to the incident laser. And hybrid coupling of those modes significantly improves the PL signals and also lightens the PL images of monolayer MoS2. This work provides a unique way to improve the emission of monolayer TMDCs. The atomically thin TMDCs coupled to plasmonic metamaterials are also promising for advanced applications such as ultrathin integrated light-emission diodes, photodetection, and nanolasers.

Strong Localization of Surface Plasmon Polaritons with Engineered Disorder.

In this work, we experimentally demonstrate for the first time strong localization of surface plasmon polaritons (SPPs) at visible regime in metallic nanogratings with short-range correlated disorder. By increasing the degree of disorder, the confinement of SPPs is significantly enhanced, and the effective SPP propagation length dramatically shrinks. Strong localization of SPPs eventually emerges at visible regime, which is verified by the exponentially decayed fields and the vanishing autocorrelation function of the SPPs. Physically, the short-range correlated disorder induces strong interference among multiple scattered SPPs and provides an adequate fluctuation to effective permittivity, which leads to the localization effect. Our study demonstrates a unique opportunity for disorder engineering to manipulate light on nanoscale and may achieve various applications in random nanolasing, solar energy, and strong light-matter interactions.

Broadband integrated polarization rotator using three-layer metallic grating structures.

In this work, we demonstrate broadband integrated polarization rotator (IPR) with a series of three-layer rotating metallic grating structures. This transmissive optical IPR can conveniently rotate the polarization of linearly polarized light to any desired directions at different spatial locations with high conversion efficiency, which is nearly constant for different rotation angles. The linear polarization rotation originates from multi-wave interference in the three-layer grating structure. We anticipate that this type of IPR will find wide applications in analytical chemistry, biology, communication technology, imaging, etc.

Influences of dietary lipid and temperature on growth, fat deposition and lipoprotein lipase expression in darkbarbel catfish (Pelteobagrus vachellii).

Darkbarbel catfish (Pelteobagrus vachellii) is an important freshwater fish in China. Water temperature greatly influences the absorption and utilization of dietary lipid by fish. Response values (including growth, hepatic fat deposition, and gene expression) for darkbarbel catfish mediated by two factors (water temperature 20-34°C; dietary lipid level 2-17%) were the focus of this study. The relationship between the two factors and the response values was evaluated by the response surface method using the central composite design. The experiment was conducted under laboratory conditions and lasted for seven weeks. A total of 975 experimental fish (average weight 11.75 ± 0.17g) were selected and placed in 39 plastic tanks. The results showed that the linear effects of lipid level on feed conversion rate (FCR), hepatopancreas somatic index (HSI), hepatic triglycerides (TG), cholesterol (TC), and lipoprotein lipase (LPL) gene expression were significant (P < 0.05). The linear effects of water temperature on specific growth rate (SGR), HSI, TC level, and LPL mRNA expression were significant (P < 0.05). The quadratic effects of water temperature and lipid level on SGR and FCR were significant (P < 0.05). Low water temperature and low lipid diets significantly inhibited growth, increased HSI, and reduced hepatic TG and TC levels, and LPL mRNA expression. The adjusted R2 values for the SGR, FCR, HSI, TC, TG, and LPL mRNA regression models were 0.77, 0.85, 0.62, 0.73, 0.85, and 0.91, respectively. The optimal combination of water temperature and dietary lipid level was 27.5°C and 9.2%, at which the greatest growth and FCR were 2.13%.d-1 and 1.31 respectively, with desirability of 0.904. These results indicated that water temperature may mediate the requirement and utilization of dietary lipid, and intervene in hepatic fat deposition. The results of this study can be used to help optimize the culture conditions of darkbarbel catfish.

Polarization-dependent strong coupling between surface plasmon polaritons and excitons in an organic-dye-doped nanostructure.

In this work, we demonstrate polarization-dependent strong coupling between surface plasmon polaritons (SPPs) and excitons in the J-aggregates-attached aperture array. It is shown that the excitons strongly couple with the polarization-dependent SPPs, and Rabi splittings are consequently observed. As a result, the polarization-dependent polariton bands are generated in the system. Increasing the incident angle, the polaritons disperse to higher energies under transverse-electric illumination, while the polaritons disperse to lower energies under transverse-magnetic illumination. Therefore, at different polarization incidence, we experimentally achieve distinct polaritons with opposite dispersion directions. In this way, tuning the polarization of the incident light, we can excite different polaritons whose energy propagates to different directions. Furthermore, by retrieving the mixing fractions of the components in these polariton bands, we find that the dispersion properties of the polaritons are inherited from both the SPPs and the excitons. Our investigation may inspire related studies on tunable photon-exciton interactions and achieve some potential applications on active polariton devices.

Dielectric lens guides in-plane propagation of surface plasmon polaritons.

In this work, we present in-plane propagation of surface plasmon polaritons (SPPs) guided by a single dielectric (Al2O3) subwavelength lens. By mounting a designed Al2O3 nanoparticle on the silver film, the effective index of a silver-Al2O3 interface is influenced by the particle thickness, then the phase difference between the silver-air and silver-Al2O3 interface can be utilized to modulate the in-plane propagation of SPPs. We show that an elliptical Al2O3 lens transforms the diffusive SPPs into a collimated beam, whose direction of propagation and beam width can be easily controlled. We also present that a triangular Al2O3 lens significantly reforms the SPPs to a Bessel beam, which possesses non-diffractive and self-healing properties. Our investigation provides unique way to guide the in-plane transport of SPPs by using dielectric subwavelength elements, which may achieve potential applications in plasmonic integrated circuits.

A novel micro-straw for cryopreservation of small number of human spermatozoon.

Cryopreservation of few spermatozoa is still a major challenge for male fertility preservation. This study reports use a new micro-straw (LSL straw) for freezing few spermatozoa for intracytoplasmic sperm injection (ICSI). Semen samples from 22 fertile donors were collected, and each semen sample was diluted and mixed with cryoprotectant in a ratio of 1:1, and then frozen using three different straws such as LSL straw (50-100 µl), traditional 0.25 ml and 0.5 ml straws. For freezing, all straws were fumigated with liquid nitrogen, with temperature directly reducing to -130--140°C. Sperm concentration, progressive motility, morphology, acrosome integrity, and DNA fragmentation index were evaluated before and after freezing. After freezing-thawing, LSL straw group had significantly higher percentage of sperm motility than traditional 0.25 ml and 0.5 ml straw groups (38.5% vs 27.4% and 25.6%, P < 0.003). Sperm motility and acrosomal integrity after freezing-thawing were significantly lower than that of before freezing. However, there was no significant difference in morphology, acrosome, and DNA integrity between the three types of straws (P > 0.05). As LSL straws were thinner and hold very small volume, the freezing rate of LSL straw was obviously faster than 0.25 ml straw and 0.5 ml straws. In conclusion, LSL micro-straws may be useful to store few motile spermatozoa with good recovery of motility for patients undergoing ICSI treatment.

Multimode photon-exciton coupling in an organic-dye-attached photonic quasicrystal.

In this Letter, we present hybrid strong coupling between multiple photonic modes and excitons in an organic-dye-attached photonic quasicrystal. The excitons effectively interact with the photonic modes offered by the photonic quasicrystal, and multiple hybrid polariton bands are demonstrated in both experiments and calculations. Furthermore, by retrieving the measured dispersion map, we get the mixing fractions of photonic modes and excitons, and show that the polariton bands inherit not only the energy dispersion features, but also the damping behaviors from both the photonic modes and the excitons. Our investigation may inspire related studies on multimode light-matter interactions and achieve some potential applications for multimode sensors.

Heterodinuclear M(II)-Ln(III) single molecule magnets constructed from exchange-coupled single ion magnets.

The synthesis and characterization of four dinuclear 3d-4f complexes [M(II)Ln(III)(L)(DBM)3] (ZnDy = 1, CoY = 2, CoDy = 3·3.5CH3CN, CoGd = 4·3.5CH3CN) are reported (H2L = N,N'-dimethyl-N,N'-(2-hydroxy-3-methoxy-5-methyl-benzyl)ethylenediamine, DBM(-) = anion of 1,3-diphenyl-propane-1,3-dione). In each of the four complexes, the M(II) ion occupies the internal N2O2 site whereas the Ln(III) ion occupies the external O4 site. There are diphenoxo bridges between the M(II) and Ln(III) ions in these complexes. The remaining coordination sites are occupied by three DBM(-) anions. Direct current (dc) magnetic susceptibility measurements indicate the presence of intramolecular ferromagnetic interactions in complexes 3 and 4. The magnetic coupling constant, JCoGd, of complex 4 is estimated to be 0.26 cm(-1) (H = -2JCoGdSCoSGd). Alternating current (ac) magnetic susceptibility studies reveal that complexes 1 and 2 show field-induced single molecule magnet behavior, with ΔE values of 36.5 K and 8.56 K, respectively. Complex 3 shows frequency dependent out-of-phase signals, indicating the presence of a slow relaxation of the magnetization, whereas complex 4 does not display slow magnetization relaxation.

[Post-thaw sperm functions of high- and low-fertility sperm donors: a comparative study of 40 cases].

OBJECTIVE: To investigate sperm function indexes that can be used to effectively evaluate the sperm donors' fertility so as to select healthy post-thaw semen samples and improve the success rate of assisted reproductive technology. METHODS: According to the pregnancy outcomes, we divided 40 donor semen samples into a high-fertility group (n = 20) and a low-fertility group (n = 20). We measured and compared the concentration, progressive motility, morphology, acrosome intactness, DNA integrity and mitochondrial membrane potential (MMP) of the post-thaw sperm between the two groups. RESULTS: There were statistically significant differences between the high- and low-fertility groups in the percentages of morphologically normal sperm ([18.50 +/- 6.10]% vs [14.42 +/- 6.44]%, P < 0.01), acrosome intactness ([86.17 +/- 4.49]% vs [80.04 +/- 7.52]%, P < 0.05) and DNA fragmentation index ([9.21 +/- 3.22]% vs [15.72 +/- 8.20]%, P < 0.05), but not in MMP ([56.75 +/- 18.80]% vs [52.23 +/- 18.86]%, P > 0.05). A significantly positive correlation was found between MMP and sperm motility (r = 0.760, P < 0.05), but not between other sperm functions and sperm concentration and motility. CONCLUSION: Sperm concentration, motility, morphology, acrosome intactness rate and DNA integrity contribute effectively to the evaluation of the fertilization capacity of post-thaw donor semen samples.

In Situ Formation of Semichelating Ligands: A Strategy for Tuning the Magnetic Coupling in Azide-Bridged Copper(II) Complexes.

Five CuII complexes of trinuclear [Cu3 (L1 )2 (N3 )6 ] (1), one-dimensional chain [Cu5 (L2 )2 (N3 )10 ]n  (2), trinuclear [Cu3 (L3 )2 (N3 )6 ] (3) and dinuclear [Cu2 (L4 )2 (N3 )2 Cl2 ] (4) and [Cu2 (L5 )2 (N3 )2 Cl2 ] (5) (L1-5 =RCH2 OR', R=substituted pyrazole or imidazole, R'=Me, Et or nPr) were synthesized by the reaction of CuCl2 ⋅2 H2 O, sodium azide with RCH2 Cl in CH3 OH, C2 H5 OH or n-C3 H7 OH. The ether ligands L1 and L2 can be alternatively prepared by the alcoholysis of RCH2 Cl in the presence of sodium azide, whereas the ligands L3-5 form in corresponding alcohols without azide. For complexes 1-5, the oxygen atom of the ether ligands is located at the Jahn-Teller axis of CuII with the long CuO separations of 2.377(3)-2.830(3) Å. The nitrogen atoms of bridging azides are located in the equatorial or basal planes of CuII ions, favoring strong magnetic coupling. The Cu-Nazido -Cu bridging angles in complexes 1-3 are 98.8(2)°-101.6(1)°, leading to ferromagnetic coupling, whereas the Cu-Nazido -Cu angles in complexes 4 and 5 are close to 103°, leading to antiferromagnetic coupling.

[Reference values of semen parameters for normal fertile men in Shanghai].

OBJECTIVE: To analyze the distribution characteristics of the main semen parameters of healthy semen donors and normal fertile men in Shanghai, compare the semen quality between the two groups, and investigate the normal reference values of the semen parameters of the fertile population in Shanghai. METHODS: We obtained semen samples from 100 healthy donors and 41 fertile men, performed semen analyses according to the WHO (2010) guidelines, and determined the semen volume, sperm concentration, sperm progressive motility, total sperm count and total progressively motile sperm count. We analyzed the distribution of the semen parameters of the normal fertile men, and obtained the lower limits of their normal reference values. RESULTS: There were no statistically significant differences in the main semen parameters between the healthy donors and normal fertile men (P < 0.05). The lower reference limits for the semen parameters of normal fertile men in Shanghai (P < 0.05) were as follows: sperm concentration > or = 27.3 x 10(6)/ml, sperm progressive motility > or = 8.1%, semen volume > or = 0.82 ml, total sperm count > or = 44.73 x 10(6) per ejaculate, and total progressively motile sperm count > or = 24.68 x 10(6) per ejaculate. CONCLUSION: For the evaluation of male fecundity, total sperm count and total progressively motile sperm count may be two better predictors than others.

[Direct fumigation for freeze-thawing of human sperm: an experimental study].

OBJECTIVE: To study the effect of direct fumigation on the post-thaw recovery rate of cryopreserved spermatozoa, and to search for a best method for human sperm cryopreservation. METHODS: We collected semen samples from 100 donors conforming to the normal reference values in WHO Laboratory Manual for the Examination and Processing of Human Semen (5th Ed), divided them into two groups, and subjected them to cryopreservation by programmable freezing (Group A) and direct fumigation (Group B), respectively. We detected the progressive motility of pre-freezing and post-thaw sperm with a computer-assisted semen analyzer, and compared the effects of the two methods on the functional integrity of sperm membrane and the rate of abnormal sperm using the percentage of hypo-osmotic swelling sperm and modified Papanicolaou staining. RESULTS: Statistically significant differences were found in post-thaw sperm progressive motility between the Groups A and B ([34.0 +/- 18.4]% vs [43.0 +/- 19.5]%, P<0.05), both remarkably decreased as compared with pre-freezing ([57.0 +/- 16.7]%, P<0.05). Such differences were also found in the post-thaw recovery rate of progressively motile sperm between the two groups ([52.2 +/- 20.6]% vs [67.1 +/- 20.0]%, P<0.05). The post-thaw percentage of hypo-osmotic swelling sperm was obviously decreased in both Groups A and B ([67.1 +/- 11.1]% and [70.6 +/- 10.0]%) in comparison with pre-freezing ([84.5 +/- 7.5]%, P<0.05), with significant differences between A and B (P<0.05). However, the rate of sperm abnormality was evidently increased in Groups A and B ([85.0 +/- 8.7% and [85.7 +/- 9.1]%), significantly higher than pre-freezing ([77.8 +/- 9.6]%, P<0.05), but with no significant differences between A and B (P>0.05). CONCLUSION: Direct fumigation is superior to programmable freezing for its easier operation, wider application, and higher sperm recovery rate.