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Most of the malignant melanomas are already in the middle and advanced stages when they are diagnosed, which is often accompanied by the metastasis and spread of other organs.Besides, the prognosis of patients is bleak. The characteristics of the local immune microenvironment in metastatic melanoma have important implications for both tumor progression and tumor treatment. In this study, data on patients with metastatic melanoma from the TCGA and GEO datasets were selected for immune, stromal, and estimate scores, and overlapping differentially expressed genes (DEGs) were screened. A nine-IRGs prognostic model (ALOX5AP, ARHGAP15, CCL8, FCER1G, GBP4, HCK, MMP9, RARRES2 and TRIM22) was established by univariate COX regression, LASSO and multivariate COX regression. Receiver operating characteristic (ROC) curves were used to test the predictive accuracy of the model. Immune infiltration was analyzed by using CIBERSORT, Xcell and ssGSEA in high-risk and low-risk groups. The immune infiltration of the high-risk group was significantly lower than that of the low-risk group. Immune checkpoint analysis revealed that the expression of PDCD1, CTLA4, TIGIT, CD274, HAVR2 and LAG3 were significantly different in groups with different levels of risk scores. WGCNA analysis found that the yellow-green module contained seven genes (ALOX5AP, FCER1G, GBP4, HCK, MMP9, RARRES2 and TRIM22) from the nine-IRG prognostic model, of which the yellow-green module had the highest correlation with risk scores. The results of GO and KEGG suggested that the genes in the yellow-green module were mainly enriched in immune-related biological processes. Finally, we analyzed the prognostic ability and expression characteristics of ALOX5AP, ARHGAP15, CCL8, FCER1G, GBP4, HCK, MMP9, RARRES2 and TRIM22 in metastatic melanoma. Overall, a prognostic model for metastatic melanoma based on the characteristics of the tumor immune microenvironment was established, which was helpful for further studies.It could function well in helping people to understand the characteristics of the immune microenvironment in metastatic melanoma and to find possible therapeutic targets.

We introduce a method which based on Bell inequalities, to study quantum phase transitions. By using the non-linear programming, we compare two different kinds of Bell inequalities, the original Bell inequality and Clauser-Horne-Shimony-Holt (CHSH) inequality. And we find that the original Bell inequality is more accurate in detecting the Bell non-locality. By defining the maximal violation of Bell inequalities, we calculate two kinds of transitions, the one is magnetic transition in the spin- $\frac {1}{2}$ XX model and the other is topological transition in the Kitaev honeycomb model. The critical points are detected successfully. Compared with traditional methods, our method requires no prior knowledge of order parameters and it is base-free.

Citrus sinensis (L.) Osbeck seedlings were fertigated with nutrient solution containing 2 [magnesium (Mg)-sufficiency] or 0 mM (Mg-deficiency) Mg(NO3)2 for 16 weeks. Thereafter, RNA-Seq was used to investigate Mg-deficiency-responsive genes in the veins of upper and lower leaves in order to understand the molecular mechanisms for Mg-deficiency-induced vein lignification, enlargement and cracking, which appeared only in the lower leaves. In this study, 3065 upregulated and 1220 downregulated, and 1390 upregulated and 375 downregulated genes were identified in Mg-deficiency veins of lower leaves (MDVLL) vs Mg-sufficiency veins of lower leaves (MSVLL) and Mg-deficiency veins of upper leaves (MDVUL) vs Mg-sufficiency veins of upper leaves (MSVUL), respectively. There were 1473 common differentially expressed genes (DEGs) between MDVLL vs MSVLL and MDVUL vs MSVUL, 1463 of which displayed the same expression trend. Magnesium-deficiency-induced lignification, enlargement and cracking in veins of lower leaves might be related to the following factors: (i) numerous transciption factors and genes involved in lignin biosynthesis pathways, regulation of cell cycle and cell wall metabolism were upregulated; and (ii) reactive oxygen species, phytohormone and cell wall integrity signalings were activated. Conjoint analysis of proteome and transcriptome indicated that there were 287 and 56 common elements between DEGs and differentially abundant proteins (DAPs) identified in MDVLL vs MSVLL and MDVUL vs MSVUL, respectively, and that among these common elements, the abundances of 198 and 55 DAPs matched well with the transcript levels of the corresponding DEGs in MDVLL vs MSVLL and MDVUL vs MSVUL, respectively, indicating the existence of concordances between protein and transcript levels.

Copper (Cu)-toxic effects on Citrus grandis growth and Cu uptake, as well as gene expression and physiological parameters in leaves were investigated. Using RNA-Seq, 715 upregulated and 573 downregulated genes were identified in leaves of C. grandis seedlings exposed to Cu-toxicity (LCGSEC). Cu-toxicity altered the expression of 52 genes related to cell wall metabolism, thus impairing cell wall metabolism and lowering leaf growth. Cu-toxicity downregulated the expression of photosynthetic electron transport-related genes, thus reducing CO2 assimilation. Some genes involved in thermal energy dissipation, photorespiration, reactive oxygen species scavenging and cell redox homeostasis and some antioxidants (reduced glutathione, phytochelatins, metallothioneins, l-tryptophan and total phenolics) were upregulated in LCGSEC, but they could not protect LCGSEC from oxidative damage. Several adaptive responses might occur in LCGSEC. LCGSEC displayed both enhanced capacities to maintain homeostasis of Cu via reducing Cu uptake by leaves and preventing release of vacuolar Cu into the cytoplasm, and to improve internal detoxification of Cu by accumulating Cu chelators (lignin, reduced glutathione, phytochelatins, metallothioneins, l-tryptophan and total phenolics). The capacities to maintain both energy homeostasis and Ca homeostasis might be upregulated in LCGSEC. Cu-toxicity increased abscisates (auxins) level, thus stimulating stomatal closure and lowering water loss (enhancing water use efficiency and photosynthesis).

We investigate how the correlated actions of quantum channels affect the robustness of entangled states. We consider the Bell-like state and random two-qubit pure states in the correlated depolarizing, bit flip, bit-phase flip, and phase flip channels. It is found that the robustness of two-qubit pure states can be noticeably enhanced due to the correlations between consecutive actions of these noisy channels, and the Bell-like state is always the most robust one. We also consider the robustness of three-qubit pure states in correlated noisy channels. For the correlated bit flip and phase flip channels, the result shows that although the most robust and most fragile states are locally unitary equivalent, they exhibit different robustness in different correlated channels, and the effect of channel correlations on them is also significantly different. However, for the correlated depolarizing and bit-phase flip channels, the robustness of two special three-qubit pure states is exactly the same. Moreover, compared with the random three-qubit pure states, they are neither the most robust states nor the most fragile states.

This present study examined excess copper (Cu) effects on seedling growth, leaf Cu concentration, gas exchange, and protein profiles identified by a two-dimensional electrophoresis (2-DE) based mass spectrometry (MS) approach after Citrus sinensis and Citrus grandis seedlings were treated for six months with 0.5 (control), 200, 300, or 400 &mu, M CuCl2. Forty-one and 37 differentially abundant protein (DAP) spots were identified in Cu-treated C. grandis and C. sinensis leaves, respectively, including some novel DAPs that were not reported in leaves and/or roots. Most of these DAPs were identified only in C. grandis or C. sinensis leaves. More DAPs increased in abundances than DAPs decreased in abundances were observed in Cu-treated C. grandis leaves, but the opposite was true in Cu-treated C. sinensis leaves. Over 50% of DAPs were associated with photosynthesis, carbohydrate, and energy metabolism. Cu-toxicity-induced reduction in leaf CO2 assimilation might be caused by decreased abundances of proteins related to photosynthetic electron transport chain (PETC) and CO2 assimilation. Cu-effects on PETC were more pronounced in C. sinensis leaves than in C. grandis leaves. DAPs related to antioxidation and detoxification, protein folding and assembly (viz., chaperones and folding catalysts), and signal transduction might be involved in Citrus Cu-toxicity and Cu-tolerance.

Low pH and aluminum (Al)-toxicity often coexist in acidic soils. Citrus sinensis seedlings were treated with nutrient solution at a pH of 2.5, 3.0, 3.5 or 4.0 and an Al concentration of 0 or 1 mM for 18 weeks. Thereafter, malate, citrate, isocitrate, acid-metabolizing enzymes, and nonstructural carbohydrates in roots and leaves, and release of malate and citrate from roots were measured. Al concentration in roots and leaves increased under Al-toxicity, but it declined with elevating nutrient solution pH. Al-toxicity increased the levels of glucose, fructose, sucrose and total soluble sugars in leaves and roots at each given pH except for a similar sucrose level at pH 2.5–3.0, but it reduced or did not alter the levels of starch and total nonstructural carbohydrates (TNC) in leaves and roots with the exception that Al improved TNC level in roots at pH 4.0. Levels of nonstructural carbohydrates in roots and leaves rose with reducing pH with a few exceptions with or without Al-toxicity. A potential model for the possible role of root organic acid (OA) metabolism (anions) in C. sinensis Al-tolerance was proposed. With Al-toxicity, the elevated pH upregulated the OA metabolism, and increased the flow of carbon to OA metabolism, and the accumulation of malate and citrate in roots and subsequent release of them, thus reducing root and leaf Al and hence eliminating Al-toxicity. Without Al-toxicity, low pH stimulated the exudation of malate and citrate, an adaptive response of Citrus to low pH. The interactive effects of pH and pH on OA metabolism were different between roots and leaves.

Scaffolds material is the key factor for bone tissue engineering, and construction of the scaffolds is also an important part. Adopting the biocompatible, biodegradable, hydroxyapatite (HAP) and sodium alginate (SA) as the molding material, using three-dimensional printing technology, choosing cross grid filling paths, we manufactured the artificial bones through self-developed 3D printing equipment. Then we measured and analyzed important parameters of the work, and did composite culture experiment. It can be seen that the prepared artificial bone scaffold has good biocompatibility. The paper provides a reference for the study of bone tissue engineering materials.

The definitions of strong superadditive deficit for relative entropy coherence and monogamy deficit of measurement-dependent global quantum discord are proposed. The equivalence between them is proved, which provides a useful criterion for the validity of the strong superadditive inequality of relative entropy coherence. In addition, the strong superadditive deficit of relative entropy coherence is proved to be greater than or equal to zero under the condition that bipartite measurement-dependent global quantum discord (GQD) does not increase under the discarding of subsystems. Using the Monte Carlo method, it is shown that both the strong superadditive inequality of relative entropy coherence and the monogamy inequality of measurement-dependent GQD are established under general circumstances. The bipartite measurement-dependent GQD does not increase under the discarding of subsystems. The multipartite situation is also discussed in detail.

Adoptive therapy using T cells modified with tumour antigen-specific T cell receptor (TCR) genes has become a popular area of research in tumour biotherapy research. However, the efficiency of this treatment is low. To increase the efficiency of this therapy, the antigen specific TCR expression in the T cells needs to be improved. Adenoviral vector-mediated gene expression is an attractive approach to bypass the issue of TCR gene modification. The efficiency of adenovirus vector serotype 5 (Ad5) infection is low due to the absence of coxsackievirus B-adenovirus receptor (CAR) expression in T cells. In the present study, a chimeric adenoviral vector (Ad5F35L) was generated; this construct contained both the natural long-shaft of Ad5 and the Ad35 knob. A transduction study showed that the Ad5F35L vector exhibited a higher transduction efficiency in human primary T lymphocytes than the Ad5 vector and the Ad5F35S vector, which contained the Ad35 natural short-shaft and the Ad35 knob. Similar transduction efficiencies were observed for both CD4(+) T lymphocytes and CD8(+) T lymphocytes and the transfection was independent of the expression of cell surface receptors. The activation of T lymphocytes resulted in an improvement of the Ad5F35L transduction efficiency in CD4(+) T cells and a decrease in Ad5F35L transduction efficiency in CD8(+) T cells. The results demonstrate that Ad5F35L is a promising viral vector and will facilitate the clinical application of tumour antigen-specific TCR gene therapy.

Background Radiotherapy is an efficient remedy in the treatment of hepatocellular carcinoma (Hca); however, some cancer cells can still survive from the radiation and the therapeutic effect is to be improved. Regulatory T cell (Tregs)–induced tumor tolerance and Akt expression play important roles in the tumor survival. This study aims to elucidate the role of radiation induces Akt expression in Tregs. Methods A rat Hca model was developed. Hca tissue was collected from the rats with or without radiotherapy. The frequency of Treg and apoptotic Treg in Hca tissue was assessed by flow cytometry. A cell culture model was used to investigate the mechanism by which the tumor-infiltrating Tregs survive from irradiation. Results After radiotherapy, the frequency of Treg was increased in Hca, the frequency of apoptotic Tregs was decreased and the expression of Akt was increased in the remaining Tregs. The results were reproduced in vitro with a cell culture model. The addition of Akt inhibitor blocked the irradiation-induced Treg survival. Tregs with high levels of Akt still preserve the immune suppressor function. Conclusions Akt plays an important role in the radiation-induced tumor-infiltrating Treg survival in Hca.

Modern services are becoming polarized. With the emergence of more and more automatic terminal services, modern services are also gradually becoming unmanned. The automatic medicine vending machine exactly emerges under the background mentioned above. The emergence of automatic medicine vending machine can reflect the reform of China’s existing medical system to a certain extent, solve the tense supply-demand relationship between doctors and patients in China, and help some patients to achieve self-improvement and self-healing without going to the hospital. For that reason, based on the principle of ergonomics, this paper will probe into and analyze how to design more humanized automatic medicine vending machine to better fit people’s practical requirement.

The method combining the construction survey and the numerical simulation is proposed in the paper to accurately obtain the deformation of the slurry shield construction under across and parallel existing tunnel. The surface settlements of the engineering construction initial position are surveyed. The reliability of the three dimensional finite element models is verified by comparing with the survey results. The deformation variation of the existing tunnel and ground surface are obtained by the elastoplasticity nonlinear calculations, showing the feasibility and effectiveness of the proposed method and the credibility and accuracy of the calculated results. Meanwhile, the measures of control deformation are presented. The surveyed settlements after the construction process of implementing measures are smaller than those calculated, indicating the effect obviously of the measures which has practical significance.

Objective: To investigate effects of bFGF and collagen on tissue-engineered cartilage in pellet culture. Methods: Rabbit rib growth-plate chondrocytes were incubated in pellet culture. bFGF and Collagen was applied alone and in combination based on the results of previous search . The histological structure of neocartilage was observed by species stained with H＆E and Immunohistochemical analysis. The DNA content of neocartilage was measured by hoechst33258 dye. The production of matrix was estimated from the determination of hydroxyproline content and Alcian Blue method. Results: Chondrocytes formed cartilage-like tissue with distinct differentiated zones and regions. The DNA content of neocartilage was enhanced by bFGF and collagen. The production of collagen and PG was promoted by collagen. bFGF and collagen could suppress the expression of collagenⅠwhile they could decelerate the decrease of collagenⅡ. The combination of bFGF and collagen had synergistic effects. Conclusion: Pellet culture could maintain chondrocytes phenotype and enhance matrix production; bFGF could stimulate the proliferation of chondrocytes, collagen could stimulate the proliferation and the matrix production of chondrocytes, it also had a positive effect on the maintenance of cell phenotype, the combination of bFGF and collagen had synergistic effects on the maintenance of chondrocytes phenotype and the regeneration of cartilage.

recent accidents of electric power cut in many south towns caused by snowstorm show the significance of electric power system security. High voltage cable terminal plays a very important part in the electric power supply system. Rubber stress cone is the key component of high voltage cable terminal, and its property is the main question in making high voltage cable terminal operates safely. In this paper, the numerical simulation model of the rubber stress cone is proposed concerning different type of rubber materials and different taper angles of rubber stress cone. Then, nonlinear finite element equations are established with increment analysis method basing on the Rivlin strain energy model of rubber material. The displacement and stress distribution of rubber stress cone are calculated by Newton-Raphson iterative method. This paper gives comparisons between different insulation rubber materials and different taper cones, whose stress distribution curves are analyzed. Thus suggestions on application of rubber insulations and the stress cone taper angle are presented. The paper also provides theoretical references to ensuring high voltage cable terminal operates safely and improving its capability to resist sleet and frost disasters.

To investigate the value of Tei index and the sensitivity of left versus right ventricular Tei index in evaluating the fetal cardiac function in pregnancy-induced hypertension syndrome in the third trimester.Fetal echocardiograms were performed in 30 women with pregnancy-induced hypertension (PIH) syndrome and 55 with normal pregnancy of the third trimester. Tei index was obtained by calculating the ratio of the isovolumic time (isovolumic contraction and relaxation time) to the ejection time of the left and right ventricle. Comparisons of the Tei index were made between the PIH group and control group, and also between the left and right ventricles in each group.Significant difference was found in the left and right ventricular Tei index between PIH group and control group. No difference was noted between the left and right ventricular Tei index in the PIH group.Tei index is a useful indicator in evaluating fetal global cardiac function, for which purpose the left ventricular Tei index can be as sensitive as the right ventricular Tei index.

To explore the feasibility of using an intracavitary convex array probe for detecting the distal extracranial internal carotid artery (ICA) by transoral carotid ultrasonography (TOCU).Forty patients underwent examinations with bilateral ICA inspected with an intracavitary convex array probe by TOCU to observe the internal diameter, visible length, peak systolic velocity (PSV), end-diastolic velocity (EDV) and resistance index (RI).Eight of the 40 patients were excluded from the observation for the presence of carotid plaques. The examination was terminated in two patients due to sensitive throat and severe pharyngeal reflex. The rest of the patients completed the examination of the internal diameter, visible length, PSV, EDV and RI, which showed no statistically significant differences among them (P0.05).Using intracavitary convex array probe, the distal extracranial ICA disease can be diagnosed with higher accuracy.

To assess the value of an intracavitary convex array probe in detecting internal carotid artery (ICA) disease.Eighty-six carotid arteries in 43 cases were examined with intracavitary convex array probe, low-frequency convex array probe and high-frequency linear probe to collect the data including the ICA visible length, peak systolic velocity (PSV), internal diameter, blood vessel shape; common carotid artery (CCA) intimae-medial thickness (IMT), PSV, and internal diameter.Significant differences were noted in the visible length, PSV of ICA, and internal diameter detected by different frequency ultrasound probes. Intracavitary probe and high-frequency probe produced significantly different findings of the blood vessel shape.Intracavitary convex array probe has important clinical value in detecting of ICA disease.

To study the value of contrast-enhanced ultrasound with high-frequency linear-array transducer in the diagnosis of carotid stenosis.The carotid arteries of 31 patients was examined with color Doppler ultrasound (CDU), harmonic contrast ultrasound (HCU) and digital subtractive angiography (DSA), respectively. The result of DSA was used as the golden standard for evaluating the accuracy, sensitivity and specificity of CDU and HCU in the diagnosis of carotid stenosis.Satisfactory images of 62 carotid arteries were obtained from the patients using HCU with high-frequency linear-array transducer. After intravenous injection of contrast agent, densely dotted hyperechogenic signals were found in the vascular lumen, with stronger intensity than that of the vascular walls. The filling defect areas in the lumen indicated vascular stenosis. Thirty carotid stenoses were detected by DSA, and 25 by CDU. The accuracy, sensitivity and specificity of CDU was 83.3%, 82.0% and 84.9%, respectively. HCU identified 29 carotid stenoses with accuracy, sensitivity and specificity of 96.7%, 96.7% and 97.0%, respectively, significantly higher than those of CDU (P0.05).HCU with high frequency linear-array transducer has better the accuracy than CDU and may serve as a good alternative for diagnosis of carotid stenosis.

To investigate effect of tumor-specific T cell receptor gene transfection on memory T cell differentiation in vitro.TCRVbeta7.1 gene was transferred into peripheral blood mononuclear cells (PBMCs) obtained from healthy adults, and the expression of Vbeta7.1 was detected by flow cytometry before and after the transfection. Memory T cell differentiation was induced by stimulation with the hepatocarcinoma cell line BEL-7402 in vitro. The expression of surface molecules CD45RO, CD45RA and CCR7 was analyzed by flow cytometry to identify the phenotype and subsets of the memory T cells. Fluorescence-activated cell sorting was performed to detect the apoptosis of the tumor cells, and enzyme-linked immunoabsorbent assay was used to determine the production of interferon-gamma (IFN-gamma) for assessing the immune function of the memory T cells.Flow cytometry showed that TCRVbeta7.1 gene was efficiently expressed after transfection. After stimulation by the tumor cells in vitro, the expression of CD45RO in TCRVbeta7.1 gene-modified T cells increased gradually, and analysis of the coexpression of CD45RA and CCR7 revealed that the effector memory T cells constituted the majority of the differentiated memory T cells. The apoptotic rate of the tumor cells induced by the T cells increased significantly with also obviously increased INF-gamma secretion in the memory T cells.Tumor-specific TCRVbeta7.1 gene transfection can promote the differentiation of the memory T cells, the majority of which belongs to effector memory T cells that perform immune functions by inducing apoptosis and cytokine secretion.

To investigate the correlation between glycodelin and endometrial receptivity and the value of serum glycodelin detection in in vitro fertilization and embryo transfer (IVF-ET) cycles for evaluation of the outcome of pregnancy.Serum glycodelin level in women receiving IVF-ET on the day of oocyte retrieval and embryo transfer was measured by ELISA, and the condition of the endometrium was examined by transvaginal ultrasonography.There was no difference in serum glycodelin level on the day of oocyte retrieval or embryo transfer between the pregnant group and nonpregnant group, but the ratio and difference of serum glycodelin levels on the days of embryo transfer and retrieval were both higher in pregnant group than in the nonpregnant group (2.54-/+2.14 vs 0.96-/+0.51, and 0.91-/+1.22 vs -0.30-/+1.14, respectively). No correlation was found between serum glycodelin levels and endometrial thickness or echo patterns.Serum glycodelin levels on the days of retrieval and embryo transfer are not correlated to endometrial thickness or echo patterns, but their dynamic changes reflect endometrial receptivity to a certain extent, which might have prognostic value for pregnancy following IVF-ET.

To investigate the value of superparamagnetic iron oxide particles (SPIO) in magnetic resonance (MR) imaging for detecting lymph node metastasis of tumors.Twelve New Zealand rabbits were used in this study, 6 of which received unilateral intra-muscular inoculation of VX2 carcinoma cells to induce lymph node metastasis of the tumor, and the other 6 served as normal control group. MR images of the lymph nodes of the rabbits were obtained before and 12 h after subcutaneous injection of SPIO, followed by image analysis in correlation with pathological examinations.On plain MR images, normal and metastatic lymph nodes showed similar signal characteristics. After administration of SPIO, the signal intensity of both normal lymph nodes and metastatic ones remained unchanged in spin echo (SE) T1-weighted images. On SE T2-weighted images, the signal intensity of normal lymph nodes significantly decreased heterogeneously, while that of all metastatic lymph nodes remained unchanged. In gradient recalled echo (GRE) T2-weighted images, the signal intensity of normal lymph nodes decreased significantly and homogeneously, while that of 4 rabbits in metastasis group remained unchanged, with the signal intensity in the other 2 rabbits decreased heterogeneously.SPIO-enhanced MR imaging can be applied to detect lymph node metastasis of the tumors.

AIM: To investigate the relationship between the exposure of phosphatidylserine(PS) and serum-dependent opsonization in peripheral blood mononuclear cells(PBMCs), to lay a foundation for exploring the influence of physiological microenvironment on the function of phagocytes.MET HODS: PBMCs were isolated from the peripheral blood of healthy individuals using Ficoll-paque and treated by RPMI 1640 medium with or without serum after washing with PBS. The phosphatidylserine(PS) exposure was detected by annexin V staining before and after in vitro serum incubation. Malondialdehyde(MDA) in treated PBMCs was examined by spect rophotometry to evaluate the interference of lipid peroxidation. The PS exposure in different subsets of PBMCs was analyzed by flow cytometry with antiCD3, anti-CD56 and anti-HLA-DR staining. Phagocytosis was detected by FCM after incubation with bacteria expressing green fluorescent protein(GFP). RESULTS: PS+ cells significantly increased in PBMCs after serum incubation, and shortterm serum incubation immediately induced the increase of PS+ cells. Serum incubation had no influence on the membrane MDAcontent, which excluded the interference by the products of lipid peroxidation. Cell subset analysis showed that HLA-DR+ mononuclear phagocytes were the main cells exposing PS in PBMCs and significantly increased in Annexin V+ cells. Bacterial phagocytosis showed that the exposed PS was involved in the serumdependent opsonization. CONCLUSION: The exposure of PS is serumdependent and is involved in the opsonization of bacteria by PBMCs.

AIM: To evaluate the long-term effects of gonadotropin-releasing hormone (GnRH)-based vaccine on levels of GnRH antibody and testosterone, and vaccine-induced immunocastration on sexual behavior of male rats. METHODS: The rats were treated with GnRH-PE40 intraperitoneally every other day for 12 wk. GnRH antibody and testosterone level in rat blood were determined by ELISA and radioimmunoassay, respectively. Morphological changes in testes and sexual behavior of rats were evaluated. RESULTS: GnRH-PE40 induced a high production in GnRH antibody, decreased the serum testosterone level, testis atrophy and sexual function in rats. CONCLUSION: Intraperitoneal administration of GnRH-PE40 produces structural and functional castration of male rat reproductive system by inducing anti-GnRH antibody.

A soluble extract from rat skeletal muscles has been used with purified mitochondrial ATPase (F1) to develop steady states with respect to glycolytic flux, the concentrations of glycolytic intermediates and inorganic phosphate, and the concentrations and ratios of adenine nucleotides. Incubations were carried out in media resembling the ionic composition in the cell cytoplasm, in an attempt to evaluate the quantitative contributions of various effectors to the overall control mechanism under simulated in vivo conditions. The primary control reaction of glycolytic flux under the conditions studied could be identified with phosphofructokinase, followed by secondary control of the reaction catalyzed by hexokinase. Glycolytic flux was increased with increasing pH over the range 6.6–7.6, both in the absence and presence of ATPase. Without other added effectors, the glycolyzing extract maintained an ATP/ADP ratio of about 50 in the pH range 7.0–7.6, and phosphofructokinase was incompletely suppressed. Addition of increasing amounts of ATPase markedly stimulated glycolytic flux coincident with lowered steady-state ATP/ADP ratios, and decreased accumulation of hexose monophosphates. Control of flux by the ATP/ADP ratio (and simultaneously altered AMP concentration) was less effective if pH (7.3 to 7.6) or phosphate concentration (2 to 20 mm) was increased. Flux through phosphofructokinase was controlled principally when the ATP/ADP ratios were varied in the range between > 50 and 15. The inhibitory effect of citrate was evaluated. Suppression of glycolytic flux and accumulation of hexose monophosphates were dependent on incubation conditions. If the pH was 7.3 or less, and the phosphate concentration low (2 mm), flux through phosphofructokinase was significantly suppressed even at citrate concentrations less than 50 μm. Simultaneous decrease in the steady-state ATP/ADP ratio and elevation of AMP was ineffective in reversing this inhibition. At higher pH and, more dramatically, when the phosphate concentration was increased, sensitivity to citrate inhibition was markedly diminished. These data, taken together with studies of respiratory control with isolated mitochondria (E. J. Davis, and L. Lumeng, 1975), J. Biol. Chem.250, 2275–2282) strongly suggest that adenine nucleotide control of both glycolysis and respiration is exerted when the ratio of free nucleotides (not protein bound) in the cytosol is in the range of 15 to > 50. The data further suggest that citrate plays an important role in the regulation of glycolysis in muscle when the ATP/ADP ratio is high (and the phosphate concentration is correspondingly low), but that this inhibition is overcome by liberation of inorganic phosphate during muscle contraction.