Search

The presence of small amounts of human serum albumin (HSA) in urine or microalbuminuria (30-300 µg/mL) is a valuable clinical biomarker for the early detection of chronic kidney disease (CKD). Herein, we report on the development of an inexpensive and disposable immunosensor for the sensitive, specific, and label-free detection of HSA using electrochemical impedance spectroscopy (EIS). We have utilized a simple one-step screen-printing protocol to fabricate the carbon-based three-electrode system on flexible plastic substrates. To enable efficient antibody immobilization and improved sensitivity, the carbon working electrode was sequentially modified with electropolymerized polyaniline (PANI) and electrodeposited gold nanocrystals (AuNCs). The PANI matrix serves as an interconnected nanostructured scaffold for homogeneous distribution of AuNCs and the resulting PANI/AuNCs nanocomposite synergically improved the immunosensor response. The PANI/AuNCs-modified working electrode surface was characterized using scanning electron microscopy (SEM) and the electrochemical response at each step was analyzed using EIS in a ferri/ferrocyanide redox probe solution. The normalized impedance variation during immunosensing increased linearly with HSA concentration in the range of 3-300 µg/mL and a highly repeatable response was observed for each concentration. Furthermore, the immunosensor displayed high specificity when tested using spiked sample solutions containing different concentrations of actin protein and J82 cell lysate (a complex fluid containing a multitude of interfering proteins). Consequently, these experimental results confirm the feasibility of the proposed immunosensor for early diagnosis and prognosis of CKD at the point of care.

Six new metal-organic coordination polymers (CPs) [Ni(L)(2,5-TDC)(H 2 O)] n (1), [Ni(L)(1,3-BDC)(H 2 O)] n (2), [Ni(L)(1,4-BDC)(H 2 O)] n (3), [Mn(L)(2,5-TDC)(H 2 O)] n (4), [Mn(L)(2,6-PYDC)(H 2 O)] n (5) and [Mn(L)(1,4-NDC)] n (6) were achieved by reactions of the corresponding metal salt with mixed organic ligands (L = 3,6-bis(benzimidazol-1-yl)pyridazine, 2,5-H 2 TDC = thiophene-2,5-dicarboxylic acid, 1,3-H 2 BDC = isophthalic acid, 1,4-H 2 BDC = terephthalic acid, 2,6-H 2 PYDC = pyridine-2,6-dicarboxylic acid, 1,4-H 2 NDC = naphthalene-1,4-dicarboxylic acid) under solvothermal condition. CPs 1-6 were characterized by single-crystal X-ray diffraction, IR, TG, XRD and elemental analyses. Their structures range from the intricate 3D CPs 1, 3, 4 and 6 to the 2D coordination polymer 2 and the infinite 1D chain 5. The CPs 1-4 and 6 underlying networks were classified from the topological viewpoint, disclosing the distinct sql (in 1), pcu (in 3 and 6), new topology (in 2), and dia (in 4) topological nets. Moreover, analysis of thermal stability shows that they had good thermal stability. Finally, magnetic properties of CPs 1-6 have been studied, the results showed that complex 2 had ferromagnetic coupling and complexes 1, 3-6 were antiferromagnetic., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

In this study, we have fabricated a simple disposable electrochemical immunosensor for the point of care testing of microalbuminuria, a well-known clinical biomarker for the onset of chronic kidney disease. The immunosensor is fabricated by screen-printing carbon interdigitated microelectrodes on a flexible plastic substrate and utilizes electrochemical impedance spectroscopy to enable direct and label free immunosensing by analyzing interfacial changes on the electrode surface. To improve conductivity and biocompatibility of the screen-printed electrodes, we have modified it with gold nanoparticles, which are electrodeposited using linear sweep voltammetry. To enable efficient immobilization of HSA antibodies, we have developed novel PS/Ag/ab-HSA nanoprobes (polystyrene nanoparticle core with silver nanoshells covalently conjugated to HSA antibodies), and these nanoprobes are trapped on the electrode surface using dielectrophoresis. Each immunosensor has two sensing sites corresponding to test and control to improve specificity by performing differential analysis. Immunosensing results show that the normalized impedance response is linearly dependent on albumin concentration in the clinically relevant range with good repeatability. We have also developed a portable impedance readout module that can analyze the data obtained from the immunosensor and transmit it wirelessly for cloud computing. Consequently, the developed immunosensing platform can be extended to the detection of a range of immunoreactions and shows promise for point of diagnosis and public healthcare monitoring., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objective: To study the coagulation properties the refrigerated whole blood stored at 4℃., Methods: Ten units of whole blood were obtained from healthy volunteer donors and stored at 4±2℃ for 21 days. Samples were collected on the day after donation and on days 2, 4, 6, 8, 10, 14 and 21 for delection including complete blood count, electrolyte, APTT, PT, Fg, blood coagulation factors, and thromboelastography(TEG)., Results: The levels of Hb, WBC, Plt, sodium and potassium in each sample accorded with standard of storing whole blood. The level of Hb, WBC, Plt and Na + decreased along with prolonging of storage time, while the K + level increased along with prolonging of stored time, APTT and PT prolonged along with prolonging of thored time, PT>17 min at d 21, the Fg level change was no-obvious, The level of factor Ⅴ and Ⅷ decreased more than 50 % of baseline on d 6 and 4 respectively; the levels of factor Ⅱ, Ⅶ, Ⅸ, Ⅹ, Ⅺ, Ⅻ showed decreasing trend, but their levels were less than 40 % of baseline values at d 21. TEG test showed that no abnormalily of R value was found, the abnormal valnes of K and Angle were observed at d 21, the abnormal value of MA was observed at d 14., Conclusion: The whole blood stored for 10 days possesses normal coagulation function showing important significance for treatment of hemorrhage from war injury and surgical openation of heart and chest.

Background: Parthenocarpy is an excellent agronomic trait that enables crops to set fruit in the absence of pollination and fertilization, and therefore to produce seedless fruit. Although parthenocarpy is widely recognized as a hormone-dependent process, hormone-insensitive parthenocarpy can also be observed in cucumber; however, its mechanism is poorly understood. To improve the global understanding of parthenocarpy and address the hormone-insensitive parthenocarpy shown in cucumber, we conducted a physiological and proteomic analysis of differently developed fruits., Results: Physiological analysis indicated that the natural hormone-insensitive parthenocarpy of 'EC1' has broad hormone-inhibitor resistance, and the endogenous hormones in the natural parthenocarpy (NP) fruits were stable and relatively lower than those of the non-parthenocarpic cultivar '8419 s-1.' Based on the iTRAQ technique, 683 fruit developmental proteins were identified from NP, cytokinin-induced parthenocarpic (CP), pollinated and unpollinated fruits. Gene Ontology (GO) analysis showed that proteins detected from both set and aborted fruits were involved in similar biological processes, such as cell growth, the cell cycle, cell death and communication. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that 'protein synthesis' was the major biological process that differed between fruit set and fruit abortion. Clustering analysis revealed that different protein expression patterns were involved in CP and NP fruits. Forty-one parthenocarpy-specialized DEPs (differentially expressed proteins) were screened and divided into two distinctive groups: NP-specialized proteins and CP-specialized proteins. Furthermore, qRT-PCR and western blot analysis indicated that NP-specialized proteins showed hormone- or hormone-inhibitor insensitive expression patterns in both ovaries and seedlings., Conclusions: In this study, the global molecular regulation of fruit development in cucumber was revealed at the protein level. Physiological and proteomic comparisons indicated the presence of hormone-independent parthenocarpy and suppression of fruit abortion in cucumber. The proteomic analysis suggested that hormone-independent parthenocarpy is regulated by hormone-insensitive proteins such as the NP-specialized proteins. Moreover, the regulation of fruit abortion suppression may be closely related to protein synthesis pathways.

Background: TIR1-like proteins act as auxin receptors and play essential roles in auxin-mediated plant development processes. The number of auxin receptor family members varies among species. While the functions of auxin receptor genes have been widely studied in Arabidopsis, the distinct functions of cucumber (Cucumis sativus L.) auxin receptors remains poorly understood. To further our understanding of their potential role in cucumber development, two TIR1-like genes were identified and designated CsTIR1 and CsAFB2. In the present study, tomato (Sonanum lycopersicum) was used as a model to investigate the phenotypic and molecular changes associated with the overexpression of CsTIR1 and CsAFB2., Results: Differences in the subcellular localizations of CsTIR1 and CsAFB2 were identified and both genes were actively expressed in leaf, female flower and young fruit tissues of cucumber. Moreover, CsTIR1- and CsAFB2-overexpressing lines exhibited pleotropic phenotypes ranging from leaf abnormalities to seed germination and parthenocarpic fruit compared with the wild-type plants. To further elucidate the regulation of CsTIR1 and CsAFB2, the role of the miR393/TIR1 module in regulating cucumber fruit set were investigated. Activation of miR393-mediated mRNA cleavage of CsTIR1 and CsAFB2 was revealed by qPCR and semi-qPCR, which highlighted the critical role of the miR393/TIR1 module in mediating fruit set development in cucumber., Conclusion: Our results provide new insights into the involvement of CsTIR1 and CsAFB2 in regulating various phenotype alterations, and suggest that post-transcriptional regulation of CsTIR1 and CsAFB2 mediated by miR393 is essential for cucumber fruit set initiation. Collectively, these results further clarify the roles of cucumber TIR1 homologs and miR393 in regulating fruit/seed set development and leaf morphogenesis.

The adsorption of external phosphorus (P) and the distribution of dissolved inorganic P (DIP) in the sediment interstitial water and sedimentary P forms were researched under the ignited water purification sludge (IWPS) addition (mixing and capping). The results showed that the adsorption ability of the IWPS increased obviously, compared with the water purification sludge (WPS). It indicated that the S max increased by 43.7%, the EPC 0 decreased by 69.1% and the DPS decreased by 54.4%. The contribution of P disappearance in the overlying water had almost no difference between the mixing and capping with IWPS, but it was obviously higher than the control in both cases. Under the mixing condition, the adsorption of external P by unit IWPS was 2.3 times of that under the capping condition, if the contacting probability between the IWPS and the overlying water was considered. Under the capping condition, the DIP in the sediment interstitial water (1-2 cm) was 33.17 times (average) of that under the mixing condition. It was attributed to the decrease of the dissolved oxygen penetration due to capping. Under the mixing condition, the external P tended to form Ca-P, while the percentages of NH 4 Cl-P and Fe/Al-P to Tot-P were bigger under the capping condition. It was suggested that the improvement of the sedimentary microenvironment by mixing with the IWPS was favorable for the adsorption of the external P and the immobilization of the internal P.

Distribution of different phosphorus (P) forms in the overlying water and the contribution of different algae to the P disappearance were investigated under disturbance with the addition of algae (Microcystis aeruginosa and Selenastrum capricornutum, respectively). The sediments and overlying water were taken from Meiliang Bay in Taihu Lake. The results showed that the concentrations of total P (TP), dissolved total P (DTP), dissolved inorganic P (DIP) and biavailable P (BAP) decreased with and without disturbance. The uptake of DTP and DIP by Microcystis aeruginosa was better than that of Selenastrum capricornutum under the disturbance, but it was the opposite without the disturbance. The disappearance of P in the overlying water was attributed completely to the physico-chemical adsorption of the suspended solids and the uptake of algae. But the contribution of suspended solids and algae depended on the disturbance. The contribution of Microcystis aeruginosa and Selenastrum capricornutum to DTP and DIP absorption was about 60% without disturbance. However, the value was reduced to 40% (Microcystis aeruginosa) and 25% ( Selenastrum capricornutum) under the disturbance. Under the disturbance and the action of algae, the distribution of sedimentary P forms changed. NH4 Cl-P and Ca-P release and Fe/Al-P increase were observed with and without disturbance. The decrease of NH4 Cl-P and Ca-P and the increase of Fe/Al-P were more obvious with disturbance than without disturbance. Selenastrum capricornutum was favor of the release of Ca-P and the formation of Fe/Al-P.

Sedimentary phosphorus forms were investigated to clarify the release of sedimentary phosphorus forms under the repeated disturbance with the addition of algae at different initial concentrations. The sediments and overlying water were taken from the Meiliang Bay in Taihu Lake. The results showed that the concentrations of NH₄ Cl-P and Res-P decreased, while the content of Fe/Al-P and Ca-P increased without disturbance. In addition, the Ca-P increased with the increase of the initial concentration of algae and the net increase of Ca-P increased by 48% (30 µg · L⁻¹), 66% (60 µg · L⁻¹), 74% (120 µg · L⁻¹), respectively. However, under the disturbance, the NH₄Cl-P and Res-P were significantly reduced, the Fe/Al-P increased significantly. The percentage of Fe/Al-P to Tot-P was up to 66. 2% (average of the 3 experiments with the addition of algae of 30 µg · L⁻¹, 60 µg · L⁻¹ and 120 µg L-¹), it was higher than the value (53.%, average of the 3 experiments) without the disturbance. Moreover, under the disturbance, the percentage of Ca-P to Tot-P was 24.1% (average of the 3 experiments with the addition of algae of 30 µg · L⁻¹, 60 µg⁻¹ and 120 µg · L⁻¹) and it was slightly lower than that (33.0%, average of the 3 experiments) without the disturbance. It is suggested that the coexistence of disturbance and algae facilitated the formation of Fe/Al-P, but the algae accelerated the formation of Ca-P without disturbance.

Background: Tyrosinase (TYR) is the key enzyme controlling the production of melanin. Very few papers have reported that andrographolide can inhibit melanin content., Objective: To investigate the effects of andrographolide on melanin synthesis., Methods: Cell viability, melanin content, TYR activity, transcriptional and protein expression levels of TYR family and other kinds of proteins involved in melanogenesis were measured after the treatments of andrographolide., Results: It was found that andrographolide decreased melanin content, TYR activity and transcriptional and protein expression of TYR family and microphthalmia-associated transcription factor (MITF) in B16F10 melanoma cells. Data showed andrographolide also decreased melanin content and TYR content in ultraviolet B (UVB) irradiation induced brown guinea pigs. Moreover, we found that melanin content and TYR activity were effectively inhibited in Human Epidermis Melanocyte (HEM) treated with andrographolide at the medium concentrations without apparent effect on cell viability. Results in experiments treated with MG-132 or cycloheximide (CHX) showed that andrographolide lowered the content of β-catenin in cell nucleus resulting from accelerating the degradation of β-catenin. Phosphorylation of glycogen synthase kinase 3β (GSK3β) and Akt decreased simultaneously. 6-Bromoindirubin-3'-oxime (BIO, inhibitor of GSK3β) and insulin-like growth factors-1 (IGF-1, activator of Akt) could reverse the decline of β-catenin in B16F10 cells induced by andrographolide., Conclusion: These results demonstrate that andrographolide can effectively suppress melanin content and TYR activity in B16F10 cells, HEM cells and UVB-induced brown guinea pig skin by decreasing phosphorylation of GSK3β dependent on Akt, promoting the degradation of β-catenin, inhibiting β-catenin into the nucleus and decreasing the expression of MITF and TYR family. Data indicate that andrographolide may be a potential whiting agent which can have great market in cosmetics and in clinical such as curing hyperpigmentation disorders., (Copyright © 2015 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.)

Objective: To establish an animal model of acute blunt scrotal trauma (BST) and evaluate the types of lesion by conventional ultrasonography (CUS) and contrast-enhanced ultrasonography (CEUS)., Methods: We made acute BST models in 21 healthy male New Zealand rabbits by striking 3 - 12 times the unilateral testes randomly selected with a 0. 5 kg iron ball falling freely from a 30 cm height. Then we evaluated the lesion types in the models by CUS and CEUS and verified our evaluation against pathological results., Results: Acute BST models were successfully established in all the 21 animals, including contusion in 10, hematoma in 6, and rupture in 5, all confirmed by pathology. CUS clearly manifested the morphology, internal echoes, and blood flow of the testes, but had a low rate of accurate diagnosis in testicular contusion for over 6 hours as well as in complex lesions. CEUS revealed an earlier perfusion of the contrast agent and shorter arriving time (AT) and time to peak intensity ( TP) in testicular contusion than in the control testes (P <0.05) , but showed no statistically significant difference between the two groups in the half time of descending peak intensity (P>0.05). For testicular hematoma, contrast agent clearly presented its outline and a delayed low enhancement in the surrounding tissue, with significant differences from the control in AT and TTP. In severe testis rupture, occasional outflow but no perfusion of contrast agent was observed., Conclusion: BST models can be established in rabbits by repeated strikes of the unilateral testes lesion of contrast agent was observed. with a freely falling iron ball. Simple contusion injury can be induced by less than 6 strikes, while complex injuries can be inflicted by more than 10. Combined application of CUS and CEUS can improve the accuracy of diagnosis of different types of lesion.

Reduction of noncrop habitats, intensive use of pesticides and high levels of disturbance associated with intensive crop production simplify the farming landscape and bring about a sharp decline of biodiversity. This, in turn, weakens the biological control ecosystem service provided by arthropod natural enemies. Strategic use of flowering plants to enhance plant biodiversity in a well-targeted manner can provide natural enemies with food sources and shelter to improve biological control and reduce dependence on chemical pesticides. This article reviews the nutritional value of various types of plant-derived food for natural enemies, possible adverse effects on pest management, and the practical application of flowering plants in orchards, vegetables and field crops, agricultural systems where most research has taken place. Prospects for more effective use of flowering plants to maximize biological control of insect pests in agroecosystem are good but depend up on selection of optimal plant species based on information on the ecological mechanisms by which natural enemies are selectively favored over pest species., (© 2012 Institute of Zoology, Chinese Academy of Sciences.)

Background: Human chimerism with normal phenotype derived from the fusion of two different zygotes is a rare phenomenon. We describe a case of a phenotypically normal 17-year-old diagnosed with dispermic chimerism during routine ABO blood grouping., Methods: ABO grouping, ABO genotyping, karyotyping, human leukocyte antigen (HLA) typing, and short tandem repeat (STR) analysis were performed., Results: Forward typing with anti-A and anti-B sera resulted in mixed-field agglutination of red blood cells. The mother and father were blood group O and AB, respectively. The proposita had O1, A201 and B alleles in the ABO locus; O1 was a maternal allele, while A201 and B were the paternal alleles. The proposita karyotype was 46,XX/46,XY. HLA typing revealed that the proposita had three alleles (46, 51, 54) at the HLA-B locus, with the additional allele of paternal origin. STR analysis identified three alleles for five of the 15 markers (D2S1338, TPOX, D8S1179, D19S433, and D21S11) analyzed in the proposita's blood- and skin fibroblast-derived DNA. The additional alleles of TPOX, D8S1179, and D21S11 were of paternal origin., Conclusions: The genetic findings suggest that this proposita was produced by dispermic fertilization of two identical haploid ova formed by parthenogenetic activation.

Objective: To evaluate contrast-enhanced ultrasonography (CEUS) in detecting testicular perfusion in acute testis contusion., Methods: We established the model of testis contusion in 11 healthy male New Zealand rabbits by randomly hitting one side of the scrotum under general anesthesia. We examined the bilateral scrotums of all the animals before, immediately after and at 2, 4 and 6 hours after modeling by color Doppler flow imaging (CDFI) and CEUS, and analyzed the time-intensity curve (TIC), arriving time (AT), time to peak intensity (TTP), peak intensity (PI), half time of descending peak intensity (HT) and area under the curve (AUC) in the healthy and injured testis, respectively., Results: CEUS exhibited a higher sensitivity in detecting tissue perfusion than CDFI. The mode of contrast agent perfusion in testicular contusion was fast in and slow out. There were no evident differences between the contused and the healthy testis in AT, TTP and PI before modeling. The contused testis showed significantly earlier AT and TTP, higher PI and larger AUC (P < 0.05) than the healthy one at different time points after modeling, but no statistically significant difference was found in HT (P > 0.05)., Conclusion: Accurate parameters of testicular perfusion in acute testis contusion can be quantitatively obtained by CEUS, which are of important value for the diagnosis of testis contusion.

The aim of the present study was to investigate the anti-proliferation and pro-apoptosis effect of Coix lachrymajobi L varma-yuan on acute T lymphoblast leukemia cell line Jurkat cells and its mechanism. Jurkat cells were treated with Coix lachrymajobi L varma-yuan of various concentrations (0, 0.4, 0.8, 1.6 mg/ml) for 24h. The inhibitory ratio was measured by Cell Counting Kit-8. The effects of Coix lachrymajobi L varma-yuan on apoptosis of Jurkat cells were determined by Hoechst 33258, PI and Annexin V-FITC/PI double staining. The mitochondrial membrane potential was analyzed by JC-1 staining. The results demonstrated that Coix lachrymajobi L varma-yuan inhibited the proliferation of Jurkat cells, and induced chromatin condensation and fragmentation (characteristic of apoptosis) and loss of mitochondrial membrane potential. In conclusion, Coix lachrymajobi L varma-yuan can inhibit the cell proliferation and induce the apoptosis of Jurkat cells. These effects relate to loss of mitochondrial membrane potential. These results suggest that Coix lachrymajobi L varma-yuan may be of value in treating lymphoma.

This study was purposed to investigate the effect of highway transportation on the quality of blood components so as to provide experimental basis to meet the needs of military operations. The transport condition was simulated by random vibration test. The red blood cells, leukocyte-reduced red blood cells, washed red blood cells were randomly vibrated (C Road) for 4 hours. Then, these blood components were stored in refrigerator for 15 days (4 degrees C). Six milliliters of blood were collected before vibration, after vibration, and at day 15 days of storage after vibration, respectively. The suspension was isolated. The free hemoglobin (FHb), routine hematological parameters, and biochemical indexes were determined. The results showed that FHb, lactate dehydrogenase (LDH), K(+) of red blood cells and leukocyte-reduced red blood cells did not significantly change after vibration and storage. However, FHb, LDH and K(+) of washed red blood cells increased significantly after simulated transportation (p < 0.05). The levels of these parameters at day 15 of storage after vibration were also significantly higher than those after vibration (p < 0.01). The changes of other hematological parameters were not significant in three blood components after vibration (C Road) and storage for 15 day. In conclusion, red blood cells and leukocyte-reduced red blood cells were qualified for clinic transfusion even after transportation within 4 hours for 15 day storage later, if they were kept in proper blood container and protected from damping. However, the washed red blood cells could not be used for clinic after similar transport in the military operations.

Objective: To determine the expressions of HO-2 and eNOS in the corpus cavernosum of castrated rats and investigate the correlation of HO-2 and eNOS with erectile dysfunction., Methods: Forty SD rats of 10 weeks were equally divided into 3 castrated groups (4, 8 and 12 weeks respectively after removal of bilateral testes) and a sham operation control group. All the rats were detected for serum testosterone (T), intracavernosal pressure (ICP) and mean arterial blood pressure (MAP) 2, 8 and 12 weeks after the surgery, the content of HO-2 analyzed by Western blot and the expressions of eNOS and HO-2 determined by immunohistochemistry., Results: Testosterone level and ICP/MAP ratio were markedly decreased after the castration (P < 0.05). The HO-2 expression was detected in the corpus cavernosum of all but the 12-week castrated rats, and the area under the curve was (341.50 +/- 99.70) in the 4-week castrated group, significantly less than in the 8-week (705.00 +/- 152. 74) and the control group (876 +/- 443. 36) (P < 0. 05), with no obvious difference between the latter two. eNOS was found mainly in the vascular endothelial cells of the corpus cavernosum, significantly less in the castrated (123.94 +/- 30. 23) than in the control group (421.21 +/- 125. 12)., Conclusion: Androgens might mediate the penile erectile function through HO-2 and eNOS in the corpus cavernosum of rats.

The aim of this study was to investigate the anti-proliferation and pro-apoptosis of triptolide on Jurkat cell line in acute T lymphocytic leukemia. The Jurkat cells were treated with various concentrations of triptolide (0, 1, 2, 4, 8, 16 microg/L) for 12 hours. The inhibitory ratio was measured by Cell Counting Kit-8 assay. The effects of triptolide on apoptosis of Jurkat cells were determined by DNA fragmentation (DNA ladder), Hoechst 33258, PI and Annexin V-FITC/PI double staining. The results demonstrated that triptolide inhibited the proliferation of Jurket cells. The 50% inhibitory concentration (IC(50)) was 4.0 microg/L. Chromatin condensation in the cells treated with triptolide could be seen by light microscopy. DNA electrophoresis showed evidence of nuclear fragmentation (DNA ladder). The hypoploid (sub-G(1)) population was increased after treatment with triptolide. The translocation of phosphatidylserine at the outer surface of the cell plasma membrane could be induced by triptolide. After treatment with triptolide for 12 hours, the rates of apoptotic cells were significantly increased. Moreover, these pro-apoptosis effects were in time-dependent manner. It is concluded that triptolide can inhibit the proliferation and induce the apoptosis of Jurkat cells. This study provides experimental basis for clinical use of triptolide in leukemia therapy.

Background & Objective: Serum protein fingerprinting technology can help to identify the molecular changes related to esophageal carcinogenesis. This study was to screen serum markers and establish the predictive models that may be of help to serologic diagnosis of esophageal squamous cell carcinoma (ESCC)., Methods: Serum samples were collected from 68 ESCC patients and 44 age-and sex-matched healthy subjects, and randomized into a training set (55 ESCC patients and 35 healthy subjects) and a blind testing set (13 ESCC patients and 9 healthy subjects). Serum samples were applied to immobilized metal affinity capture (IMAC3) proteinchip surfaces and tested by surface-enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS). The data were analyzed by Biomarker Wizard software to screen serum proteomic biomarkers. Decision classification tree models were established by bioinformatics. Double-blind test was used to determine the sensitivity and specificity of the classification tree models., Results: A total of 78 effective protein peaks were detected at the molecular range of 1.5 to 20 ku, among which 25 were significantly different between ESCC patients and healthy subjects (P<0.001). All the peptide pattern data were sampled randomly for 1,000 times using 3-cross validation approach, and 1,000 decision tree models were obtained. Twenty decision trees with the highest cross validation rate were chosen to construct the classification models which can differentiate ESCC patients from healthy subjects. With these decision trees, 18 samples were correctly forecasted from 22 blind testing samples, with a sensitivity of 92.31% and a specificity of 66.67%., Conclusion: SELDI-TOF-MS technique combined with decision tree model can help to identify serum proteomic biomarkers related to ESCC and the predictive models can discriminate ESCC patients from healthy people effectively.

Objective: To investigate the expression of Rho-kinase and heme oxygenase and the interaction among NOS/NO, HO/CO and RhoA/Rho-kinase in the corpus cavernosum of spontaneous hypertensive rats (SHRs)., Methods: A series of electric stimuli were applied to the corpus cavernosum nerves of 7 SHRs, the changes of ICP/MAP observed continuously and the expressions of ROCK2, HO-2 and eNOS analyzed by Western blot and immunohistochemistry. Another 7 WKY rats were taken as controls., Results: Compared with the controls, ICP/MAP did not rise obviously (P < 0.05), HO-2 dropped sharply (P = 0.006) and the expression of the ROCK2 protein was elevated markedly (P = 0.017) in the SHRs. Immunohistochemistry showed HO-2 to be distributed in the smooth muscle cells and nervous cells of the corpus cavernosum, and eNOS mainly in the vascular endothelial cells. The expressions of HO-2 and eNOS were obviously reduced in the SHRs., Conclusion: NOS/NO, HO/CO and RhoA/Rho-kinase are related with erectile dysfunction in SHRs and may interact on one another.

Erectile dysfunction (ED) has been plaguing men for a long time and the incidence of this disease is as high as 52% among males aged between 40 and 70. Recent discovery has shown a connection between the RhoA/Rho-kinase signaling system and ED. This paper reviews the progress in the study of RhoA/Rho-kinase signaling pathway, expounds its mechanism in penile erection and provides a base for further research on the role of RhoA/Rho-kinase signaling pathway in penile erection.

Background: Spondyloepiphyseal dysplasia congenita (SEDC) is an autosomal dominant skeletal dysplasia characterized by short stature, abnormal epiphyses, and flattened vertebral bodies. Secondary prevention of SEDC can be achieved by prenatal diagnosis. Reports of antenatally-diagnosed SEDC fetuses have been very rare and molecular prenatal diagnosis even rarer. We previously reported a familial G504S mutation in the type II collagen (COL2A1) gene resulting in SEDC. In this study, molecular prenatal diagnosis was performed to 2 couples in this family with pregnancies at risk for SEDC., Methods: Amniotic fluid was sampled by amniocentesis under ultrasound guidance at 19+3 and 18+6 weeks' gestation, respectively. Karyotype and molecular genetic analysis were performed on cultured amniotic fluid cells. Maternal cell contamination was excluded by short tandem repeat (STR) analysis. Direct DNA sequencing and DHPLC were conducted to detect the potential mutation in exon 23 of COL2A1 gene. Both women underwent serial sonograms because they insisted that the molecular diagnosis should be confirmed by another method, although they had been informed that mutation analysis is predictive of the disease., Results: Karyotype of both fetuses was normal and molecular genetic analysis revealed that fetus 1 carried a G504S mutation in exon 23, while fetus 2 was normal. In case 1, femur length of the fetus was markedly below the 5th centile at 23 weeks' gestation, which confirms the accuracy of molecular diagnosis. A medical termination was carried out at 27+5 weeks' gestation and a male fetus with a relatively large head and short limbs was delivered. The fetal radiograph demonstrated a number of features, including generalised platyspondyly, absent ossification of the vertebral bodies in the cervical region and significant shortening of the long bones. The diagnosis of SEDC was thus confirmed clinically. Ultrasound monitoring of fetus 2 showed that its femur length was normal for gestational age at repeated scans, which was consistent with the molecular diagnosis., Conclusions: Molecular analysis allows early and accurate prenatal diagnosis for SEDC once mutation is known in a family. However, considering the poor genotype/phenotype correlation in many cases of SEDC, the combination of ultrasound as well as molecular genetic approach might be needed.

Objective: To investigate the therapeutic potential of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF superfamily, and to analyze TRAIL-induced apoptosis in Jurkat cells., Methods: Expression of TRAIL receptors (DR4 and DR5) was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Cytotoxic effects were determined by colony formation assay and a cell counting kit. The effects of recombinant TRAIL on apoptosis of Jurkat cells were determined by DNA fragmentation (DNA ladder) and PI staining. Changes in mitochondrial membrane potential were detected with JC-1 fluorescence., Results: TRAIL inhibited the proliferation and induced internucleosomal DNA fragmentation (characteristic of apoptosis) and loss of mitochondrial membrane potential., Conclusion: Recombinant soluble TRAIL can be used as a therapy for cancer.

Objective: To observe therapeutic effect of Yuanluo Tongjing needling method for treatment of mild cognitive impairment and search for an effective therapy of TCM for mild cognitive impairment., Methods: Sixty cases were randomly divided into an observation group and a control group, 30 cases in each group. The observation group were treated by Yuanluo Tongjing needling method with Baihui (GV 20), Danzhong (CV 17), Guanyuan (CV 4), Shenmen (HT 7), etc. selected, and the control group by oral administration of Aricept, 2.5 mg. Changes of scores of all indexes in clinical memory scale and memory quotient (MQ) before and after treatment were investigated in the two groups, and clinical therapeutic effects were assessed by using MQ increasing values., Results: After treatment for 30 days, the scores of memory quotient, direction memory, picture free memory, re-cognition of no-significant picture forms and associated memory of human picture characteristics significantly increased (P < 0.01, P < 0.05) in the observation group; and all the indexes except associated memory in the control group significantly changed (P < 0.05), with no significant difference between the two groups (P > 0.05). The good rate for memory effect was 80.0% in the observation group and 76.7% in the control group with no significant difference between the two groups (P > 0.05)., Conclusion: Yuanluo Tongjing needling method can improve memory ability of the patient of mild cognitive impairment.

The purpose of study was to investigate the impact of killer immunoglobulin-like receptor (KIR) and its ligand on haploidentical bone marrow transplantation. 74 cases were analyzed for the distribution frequencies and characteristics of KIR and its ligand as well as the impact of KIR ligand for the haploidentical bone marrow transplantation in terms of the overall survival, disease-free survival (DFS), GVHD and relapse. The results showed that among the 19 KIR genotypes currently nominated KIR2DL1, KIR2DL4 and KIR3DL2-3 could be detected in all the cases. Other high frequency genotypes included KIR3DP1 (98.6%), KIR2DP1 (98.6%), KIR3DL1 (97.3%) and KIR2DL3 (97.3%). Inhibitory receptor genotypes were 1.37-fold of activating receptor genotypes. KIR2DL1, KIR3DL2, KIR3DL3 and KIR2DL4 were found in all haplotypes and at least one genotype of KIR2DL2 and/or KIR2DL3 existed in all haplotypes. Among the 14 genotypes found in the test, the HLA-Cw7 was the most popular (37.8%) and the group 2 (HLA-Cw1, 3, 7, 8, 13, 14) recognized by KIR2DL2/2DL3 counted for 43.2%. The incompatibility of KIR for 32 cases of haploidentical BMT was 43.8%, of which 9/14 were KIR2DL incompatible, 5/14 were KIR2DL2 or KIR3DL1 incompatible. Among the 46 cases of haploidentical BMT, 29 cases were HLA-Cw matched and 14 cases were mismatched. The completed mismatch ratio of HLA-Cw was 30.4% and the match ratio was 63.4%. The survival rate was higher for the 14 cases of KIR genotype compatible group than the 13 cases of KIR genotype incompatible group (p = 0.032). The disease-free survival was significantly higher for the 17 cases of mismatched KIR ligands (HLA-Cw) group than the matched group (p = 0.024). The survival rate was higher in GVHD group than that in non-GVHD group when the KIR ligand was missing. The acute and severe GVHD was related to the existence of activating receptor of KIR2DS1/2DS2. The incompatibility group was accompanied with frequent acute and severe GVHD and less relapse and vice versa for the compatibility group. One patient died after BMT among the 14 mismatched KIR ligand group suffering from myelogenous leukemia while 4 patients out of 12 patients died in the matched group. It is concluded that the haploidentical BMT is characterized by mismatch between donor and recipient and its immunological reactions also features by the incompatibility of KIR genotype and missing ligand. The missing ligand for the donor KIR has strong effect on the outcome of BMT and it means a lot to analyze the KIR genotype and its ligand for the selection of best donor and prognostic evaluation in haploidentical BMT.

This study was aimed to investigate the therapeutic effect of growth factor-primed donor peripheral mononuclear stem cell infusion (DMNCI) for patients with relapsed leukemia after haploidentical bone marrow transplantation (BMT). The donor was the same individual for both BMT and DMNCI. All the three patients described here were Philadelphia chromosome positive leukemia before haploidentical BMT; one case was newly diagnosed as acute lymhoblastic leukemia (ALL) and the others were chronic myeloid leukemia (CML). Two cases (one with ALL and one with CML) manifested with clinical relapse and the third case was in the stage of molecular relapse. The former 2 patients received a single bulk dose of DMNCI, the inoculums of which contained mononuclear cells of 8.25 x 10(8)/kg or 5.24 x 10(8)/kg and CD3-positive cells of 1.87 x 10(8)/kg or 1.14 x 10(8)/kg respectively. The third case received initial dose of DMNCI which was 2.0 x 10(7)/kg, and received CD3 positive cells of 1.1 x 10(7)/kg. The results indicated that the different therapeutic responses were found in all three patients. Two patients with clinical relapse received temporal remission, and died of severe graft versus host disease (GVHD), relapse and failure at day 41 and 49 after DMNCI. The third patient with molecular relapse received molecular remission after 2 infusions of DMNCI. All three patients developed acute GVHD, but two patients among them developed GVHD of grad IV, other one developed GVHD of grad I and has survived in disease-free state during half a year follow-up. It is concluded that the DMNCI may be effective for the treatment of relapsed leukemia after haploidentical BMT and this treatment can be safe if the initial dose of DMNCI is 10(7)/kg and subsequent single dose of DMNCI gradually increases.

Objective: To clone human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL, 114-281) cDNA and develop an inducible system for expression in E. coli., Methods: The human TRAIL (114-281) cDNA was amplified with the total RNA from the human peripheral blood monocytes by RT-PCR. The cDNA was inserted into pMD T vector. The selected integrants were confirmed by PCR screen, digestion with restriction enzymes and DNA sequencing. Then, the insert of human TRAIL cDNA was subcloned into prokaryotic expression vector pET28a. The construction of prokaryotic expression vector was proofed correct by RT-PCR and digestive identification. The recombinant protein was induced by IPTG (isopropyl-beta-D-thiogalactopyranoside) . The sTRAIL inclusion bodies were refolded by dilution method. Refolded protein was purified with column chromatography., Results: Agarose gel electrophoresis of product of RT-PCR revealed a band around 500bp, which was expected. The positive integrants were confirmed by PCR screen and digestion with restriction enzymes. The same band as RT-PCR product was showed by PCR screen and digestion with restriction enzymes. Then the selected integrants were confirmed by DNA sequencing. The sequence was checked in GenBank. The construction of prokaryotic expression vector pET 28a was proofed correct by RT-PCR and digestive identification. TRAIL protein was successfully induced by IPTG in E. coli BL21. The results also showed that the protein was expressed as inclusion bodies. After the sTRAIL inclusion bodies were solubilized and refolded, the protein expressed was purified with one band, which was about 20kD, analyzed by SDS-PAGE., Conclusion: These results suggested that the hsTRAIL was cloned, expressed and purified in the present study. Significant quantities of TRAIL produced by this method should allow further studies in determining the physiological significance and function of TRAIL in the future.

Objective: To develop a nested polymerase chain reaction (PCR) technique for fetal SRY gene identification using cell-free fetal DNA in maternal plasma., Methods: Peripheral blood samples were obtained from 30 pregnant women and cell-free DNA was extracted by the phenol/chloroform method from plasma. The nested PCR was carried out to amplify the fragment of SRY gene by two sets of PCR primer pairs. Direct sequencing analysis was then performed on the PCR product., Results: Among the 17 women bearing male fetuses, SRY sequences were detected in 15 plasma samples after nested PCR amplification, while none of the 13 women bearing female fetuses had the positive results. The accuracy and sensitivity were 93.3% (28/30) and 88.2% (15/17), respectively., Conclusion: The phenol/chloroform extraction for fetal DNA in maternal plasma was effective and simple. And the nested PCR amplification of SRY sequence is a convenient and low-cost approach for the non-invasive early prenatal diagnosis of sex-linked inheritant diseases.

The improvement of diagnosis and treatment of andrological disease depends on scientific advances in the corresponding domain of andrology. In this article, the author thought topic selection was of great significance for the improvement of diagnosis and treatment based on his clinical and scientific experience for years in the field of andrology. Topic selection should be closely combined with clinical practice and follow the principles of importance, innovation, feasibility and reality, which meets the demands of basic, therapeutic and epidemiologic aspects in andrology research. Several common approaches to topic selection were discussed as well in the article.

The precursor of prostate specific antigen (proPSA) are distinct molecular forms of free PSA in serum. proPSA is comprised of native proPSA as well as several truncated forms, in which [-2] proPSA and [-4] proPSA are more prostate cancer (PCa)-associated than [-7] proPSA and [-5] proPSA. Clinical studies have recently provided evidence that [-2] proPSA can significantly improve the detection of PCa, particularly in patients with total serum PSA values less than 4 microg/L. In this paper, the mechanism and characteristics of proPSA formation and impact of proPSA on the early detection of PCa are reviewed.

Objective: To evaluate total antioxidant capacity (TAC) of seminal plasma in fertile and infertile men and understand the relation between seminal plasma TAC and male fertility., Methods: Two hundred and twenty-five infertile men were divided into 10 cases of obstructive azoospermic men, 42 cases of non-obstructive azoospermic men,20 cases of oligozoospermic men, 78 cases of asthenozoospermic men, 57 cases of oligoasthenozoospermic men, and 18 cases of normozoospermic men, then 28 fertile men were taken as the control. The seminal parameter analysis was performed by computer-assisted semen analysis (CASA) system. Seminal plasma TAC was measured using spectroscopic analysis., Results: Seminal plasma TAC were (1.71 +/- 1.33) U in obstructive azoospermic men, (12.73 +/- 9.44) U in non-obstructive azoospermic men, (10.85 +/- 6.64) U in oligozoospermic men, (13.88 +/- 8.24) U in asthenozoospermic men, (11.20 +/- 7.02) U in oligoasthenozoospermic men, (18.07 +/- 8.73) U in normozoospermic men, and (19.82 +/- 6.33) U in fertile men. There was no significant difference in TAC between normozoospermic men and fertile men (P > 0.05). Compared with fertile men, seminal plasma TAC in other infertile groups was significantly lower (P < 0.01). There were significantly made positive correlation between seminal plasma TAC and sperm density (r = 0.182, P < 0.05), as well as sperm with grade a (r = 0.150, P < 0.05)., Conclusion: Seminal plasma TAC is closely related to male fertility, and the decreased level of TAC in seminal plasma may be one of the causes of male infertility.

Haploidentical donors are available for most patients who need allografts but do not have matched donors. However, GVHD, rejection, delayed immune reconstitution, and infections have been significant barriers. We designed a haploidentical BMT protocol focusing on prevention of GVHD and rejection. A total of 53 leukemic patients underwent haploidentical G-CSF-primed BMT without ex vivo T-cell depletion. GVHD prophylaxis consisted of antithymocyte globulin, cyclosporine, methotrexate, and mycophenolate mofetil. In all, 38 patients (the CD25 group) received additional anti-CD25 monoclonal antibody basiliximab. The results were compared to 15 patients who did not receive basiliximab. All patients achieved trilineage engraftment with full-donor chimerism. The incidence of acute II-IV GVHD was 11% in the CD25 group vs 33% in the control group (P=0.046). The overall incidence of extensive chronic GVHD was 15%. T, B, and NK cells recovered within 12 months post transplant. The disease-free survival at 2 years was 53% with a median follow-up of 31 months. In conclusion, G-CSF primed haploidentical BMT along with sequential immunosuppressive agents as described here deserves further study., (Bone Marrow Transplantation (2005) 36, 349-354.)

Objective: To investigate the significance of prostate specific antigen (PSA) examination in seminal plasma of infertile patients., Methods: Eighty-five infertile patients were collected randomly. The level of PSA in seminal plasma was detected by ELISA method. The correlations between PSA and several seminal parameters including sperm density, motility and acid phosphatase (ACP) were analyzed., Results: The PSA, ACP concentrations and sperm motility in 65 cases of abnormal liquefaction patients were obviously lower than those in normal liquefaction patients(P < 0.01). But there were no significant differences in sperm density among the three groups(P > 0.05). PSA levels were significantly correlated with ACP and sperm motility(P < 0.01)., Conclusions: The seminal PSA in infertile patients is markedly correlated with semen liquefaction. The abnormal quality and quantity of PSA can result in a depression of sperm motility and subinfertility.

Men with non-obstructive azoospermia(NOA) can now be treated by using intra-oocyte round spermatid injection(ROSI) or elongated spermatid injection(ELSI). Spermatids can be retrieved from semen or from testis biopsy specimens. But the rates of fertilization and pregnancy with spermatids have been disappointing. Many problems limiting success rate and hindering a wide application of this technique still remain unresolved, including the incomplete maturation of spermatid nuclear, oocyte activation and identification of a live spermatid.

Objectives: To investigate the possible differences in the inhibin B levels of seminal plasma and serum between fertile and infertile males and to obtain information on the relation between serum inhibin B or seminal plasma inhibin B and spermatogenesis., Methods: Semen and blood samples were collected from fertile(n = 20), oligospermia(n = 20), asthenospermia(n = 22) and non-obstructive azoospermia(NOA) (n = 20) males at 8:00 am = 10:00 am. Semen parameters were analyzed. Levels of inhibin B in seminal plasma and serum, ACP, Fru, alpha-Glu in seminal plasma, serum levels of FSH, T, LH were determined., Results: Both levels of serum inhibin B and levels of seminal plasma inhibin B correlated significantly negatively with serum FSH(r = -0.536, P < 0.001 vs r = -0.288, P = 0.01), and statistically positively with sperm concentration(r = 0.49, P < 0.001 vs r = 0.48, P < 0.001). There was positive correlation between levels of seminal plasma inhibin B and activity of alpha-Glu in seminal plasma (r = 0.377, P = 0.001). The difference in levels of seminal plasma inhibin B was found only between fertile males or asthenospermia and NOA (P < 0.01 and P < 0.05, respectively). However, significant differences in levels of serum inhibin B were found not only between males with normal sperm concentration (including fertile males and asthenospermia) and NOA (P < 0.01), fertile males and oligospermia (P < 0.05), but also between oligospermia and NOA (P < 0.05). There was no correlation between serum inhibin B and seminal plasma inhibin B., Conclusions: Both levels of serum inhibin B and seminal plasma inhibin B could reflect testis spermatogenesis status. Levels of seminal plasma inhibin B could also reflect the function of seminiferous duct, but the wide range of values limited its applicability.

Objectives: To detect the cholesterol ester transfer protein (CETP) levels in semen of infertile patients and evaluate the correlation between CETP in semial plasma and infertility., Methods: One hundred and sixty-three infertile patients and fifteen fertile males were selected randomly. The routine examination of ejaculates was fulfilled by computer aided semen analysis (CASA). The CETP levels in all seminal plasma samples and fifty-five serum samples were detected by ELISA method., Results: The CETP levels in infertile patients and fertile males were (2.21 +/- 1.23) microgram/L and (1.40 +/- 0.45) microgram/L, respectively. There were no significant differences between the two groups(P > 0.05). And there were no significant differences of CETP levels in seminal plasma among groups of azoospermia(n = 29), oligoasthenozoospermia (n = 58), oligospermia(n = 15), asthenozoospermia(n = 44) and normozoospermia(n = 17) in the infertile patients(P > 0.05). The CETP in seminal plasma and serum were detected in 55 infertile patients, and there was no correlation between CETP levels in seminal plasma and serum using Spearman analysis(r = 0.009, P > 0.05). The mean CETP level in seminal plasma was almost 1/1,000 of that in serum., Conclusions: The CETP level in seminal plasma is extremely low and has no relation with the changes of sperm density or motility. It may ensure the integrity of sperm membrane before the sperm enters into female genital tract.

Objectives: To evaluate the clinical significance of the detection of IgG and IgM antibodies against Chlamydia trachomatis (CT) in semen of asymptomatic infertile patients., Methods: One hundred and sixteen asymptomatic infertile patients and eighteen fertile males were selected randomly. The routine parameter analysis of semen was fulfilled by computer aided semen analysis(CASA). Then the seminal plasma was separated and the IgG, IgM antibodies against CT in seminal plasma were determined with ELISA method., Results: IgG and IgM antibodies against CT were present in 13.8% (16/116) and 3.4% (4/116) of the semen of infertile patients, while for the fertile males the percentages were 11.1% (2/18) and 0, respectively. There were no differences between the two groups(P > 0.05). In the infertile patients, 22 patients were azoospermia. And in the rest 94 infertile patients, the percentages of IgG and IgM antibodies in abnormal sperm density group were 21.4% (6/28) and 7.1% (2/28), which were higher than those in normal group, but there were no statistical differences(P > 0.05). Similarly, the IgG, IgM antibodies were not correlated with the sperm motility(P > 0.05). The positive percentage of CT in 116 patients was 25.9% (30/116)., Conclusions: The percentages of IgG and IgM antibodies against CT in semen of asymptomatic infertile patients are similar to that in fertile males, which do not correlate with the changes of semen parameters, and may not be used for indication of CT infection.

Objectives: To develop a method by which large and purified populations of spermatids can be isolated in semen of male infertile patients., Methods: A total of fifteen ejaculates containing cellular elements from infertile patients with various andrological pathologies were obtained after a 24-hour abstinence. A modified discontinuous Percoll gradient (15%, 22%, 30%, 40%, 50%, 60%) centrifugation method was used to isolate the spermatids. After centrifugation at 2,000 r/min for 30 min at 18 degrees C, the single Percoll fractions were separated and analyzed in order to select the one with the greatest purity of spermatid. The germinal cells in each isolated fraction were counted using a Macro sperm counting chamber, then the contents of spermatids were determined by morphology (Wright-Giemsa staining method) and flow cytometry (FCM) analysis, while the contaminated leukocytes were assessed by anti-CD45 immunocytochemistry., Results: After Percoll centrifuged, six single fractions were obtained. Morphology and FCM analysis showed that the 22% fraction contained mostly spermatids [(91.85 +/- 5.18)%, P < 0.005] and the mean density in this fraction was (1.010 +/- 0.786) x 10(5)/ml. While in the 30% fraction, various immature spermatogenic cells including spermatids were present and leukocytes mostly presented in the 60% fraction., Conclusions: A large population of relatively purified spermatids can be isolated from the ejaculates of infertile patients by using this modified discontinuous Percoll gradient centrifugation method.

Aim: To analyze the relationship between autosomal aberrations and testicular dysgenesis or spermatogenic arrest in Chinese patients and to map the corresponding regions on each autosome in regard to the recorded aberrations accompanying these distubances., Methods: One hundred and nineteen cases of aberrant karyotypes with testicular dysgenesis, azoospermia or oligozoospermia reported in five Chinese journals and one monograph were analyzed. For each autosome, the type and frequency of chromosomal aberrations were counted and the regions corresponding to the disturbances were mapped out., Results: Chromosomes 13, 14, 9, 21 exhibited a high frequency of aberration and bands 14q11 and 13p11 were the two regions showing the highest linkage to testicular dysgenesis or infertility. The frequency of chromosomal aberrations was higher in bands 9p11 and 22q than in others., Conclusion: Autosomes 13, 14, 9 and 21 in the order of importance play a critical role in testicular development and spermatogenesis and other autosomes may also contribute; the following regions, 14q11, 13p11,9p11, and 22q, are of high significance.

Objectives: To develop a simple and effective method by which spermatids can be isolated from mouse testis., Methods: Combination of enzymatic digestion was used to prepare suspension of spermatogenic cells from adult mouse testis, and then a modified discontinuous Percoll gradient (15%, 22%, 30%, 40%, 50%, 60%) centrifugation method was introduced to isolate spermatids from the cellular suspension. The content of spermatids in each isolated fraction by Percoll method was determined by morphology (Wright-Giemsa staining) and flow cytometry analysis, and the viability of spermatogenic cells was assessed using Eosin Y exclusion test., Results: More than 97% of the testicular cells remained their viability after enzymatic digestion. After Percoll centrifuged, six fractions were formed. In each isolated fraction, the 22% fraction contained mostly spermatids(mean 86.7%) and cell viability was more than 85.5%. While in the 30% fraction, immature spermatogenic cells were present, and more than 92% of the cells remained their viability., Conclusions: A large of relatively purified spermatids can be isolated from mouse testis by enzymatic digestion combined discontinuous Percoll gradient centrifugation method.