[Short-term nitrogen addition reduces soil microbial nitrogen fixation rate in subtropical Pinus taiwanensis and Castanopsis faberi forests]. / çŸ­æœŸæ°®æ·»åŠ é™ä½Žäºšçƒ­å¸¦é»„å±±æ¾æž—å’Œç½—æµ®æ ²æž—åœŸå£¤å¾®ç”Ÿç‰©å›ºæ°®é€ŸçŽ‡.

Biological nitrogen (N) fixation is an important source of N in terrestrial ecosystems, but the response of soil microbial N fixation rate to N deposition in different forest ecosystems still remains uncertain. We conducted a field N addition experiment to simulate atmosphere N deposition in subtropical Pinus taiwanensis and Castanopsis faberi forests. We set up three levels of nitrogen addition using urea as the N source: 0 (control), 40 (low N), and 80 g N·hm-2·a-1(high N) to examine the chemical properties, microbial biomass C, enzyme activities, and nifH gene copies of top soils (0-10 cm). We also measured the microbial N fixation rate using the 15N labeling method. Results showed that N addition significantly reduced the soil microbial N fixation rate in the P. taiwanensis and C. faberi forests by 29%-33% and 10%-18%, respectively. Nitrogen addition significantly reduced N-acquiring enzyme (i.e., ß-1, 4-N-acetylglucosaminidase) activity and nifH gene copies in both forest soils. There was a significant positive correlation between the microbial N fixation rate and soil dissolved organic C content in the P. taiwanensis forest, but a significant negative relationship between the rate of soil microbial nitrogen fixation and NH4+-N content in the C. faberi forest. Overall, soil microbial N fixation function in the P. taiwanensis forest was more sensitive to N addition than that in the C. faberi forest, and the factors affecting microbial N fixation varied between the two forest soils. The study could provide insights into the effects of N addition on biological N fixation in forest ecosystems, and a theoretical basis for forest management.

F-actin/DRP1 axis-mediated mitochondrial fission promotes mitophagy in diabetic submandibular glands.

BACKGROUND: Diabetes is accompanied by a high prevalence of hyposalivation, causing severe damage to oral and systemic health. Mitochondrial dynamics play important roles in the pathogenesis of various diabetic complications; however, little is known about their roles in diabetic hyposalivation. MATERIALS AND METHODS: A diabetic mouse model and a high glucose (HG)-induced diabetic submandibular gland (SMG) cell model were employed. RESULTS: More mitochondria surrounded by autophagosomes and higher expression of mitophagy-related proteins were detected in the SMGs of diabetic mice and HG-treated SMG cells. In diabetic SMGs, dynamin-related protein 1 (DRP1) was upregulated, whereas mitofusin-2 was downregulated both in vivo and in vitro. Shortened mitochondria and impaired mitochondrial functions were observed in the HG group. A DRP1-specific inhibitor, mdivi-1, suppressed mitochondrial fission and mitophagy, as well as restored mitochondrial functions in the HG condition. Moreover, the interaction of F-actin and DRP1 was enhanced in the diabetic group. Inhibiting F-actin with cytochalasin D repaired the injured effects of HG on mitochondrial dynamics and functions. Conversely, the F-actin-polymerization-inducer jasplakinolide aggravated mitochondrial fission and dysfunction. CONCLUSIONS: F-actin contributes to HG-evoked mitochondrial fission by interacting with DRP1, which induces mitophagy and impairs mitochondrial function in SMG cells, ultimately damaging the SMG.

Microwave-assisted esterification and electro-enhanced solid-phase microextraction of omega-3 polyunsaturated fatty acids in eggs.

Omega-3 polyunsaturated fatty acids (ω-3 PUFAs), a type of fatty acid that has many health benefits, are of increasing concern. Herein, we developed a method for the rapid esterification and enrichment of ω-3 PUFAs in eggs, which includes microwave-assisted esterification (MAE) and electrically enhanced solid-phase microextraction (EE-SPME). Combined with gas chromatographic, efficient detection of ω-3 PUFAs was achieved in eggs. Under microwave radiation, the esterification efficiency exhibited a significant increase ranging from 5.06 to 10.65 times. The EE-SPME method reduced extraction time from 50 to 15 min. In addition, improvements in extractive fiber coating materials were explored, which ensured efficient extraction of ω-3 PUFAs. Under the optimal conditions, the method displayed a low detection limit (1.01-1.54 µg L-1), good recoveries (85.82%-106.01%), and wide linear range (7.5-1000 µg L-1), which was successfully applied to determine ω-3 PUFAs in real egg samples.

Membrane-protected magnetic covalent organic framework for efficient extraction of estrogens in dairy products.

The presence of estrogens residues in dairy products is a growing concern due to their potential health risk. Herein, in this study, we have developed a membrane-protected magnetic solid-phase extraction (MP-MSPE) method that utilized a magnetic adsorbent (Fe3O4@COF-LZU1) with in-situ growth for the efficient extraction of estrone (E1), 17ß-estradiol (E2), and estriol (E3). When combined with HPLC-FLD, this method allows for the efficient detection of estrogens in dairy products. The stability of the MP-MSPE was improved by the presence of a dialysis membrane, which remained a high extraction efficiency (90 %) even after ten reuse cycles. The hydrogen bonding, π-π interactions and pore size effect contribute to the excellent adsorption of three estrogens onto Fe3O4@COF-LZU1. Under optimal conditions, the method exhibits a low detection limit (0.01-0.15 µg L-1), wide linear range (0.1-800 µg L-1), and favorable recoveries (77.3 %-109.4 %) at three concentration levels (10, 50 and 100 µg L-1). This proposed method is characterized by its simplicity, high efficiency and eco-friendliness, making it a promising approach for extracting estrogens from dairy products.

[Relationship between Iron Metabolic Parameters and Platelet Counts in Blood Donors].

OBJECTIVE: To investigate the correlation of iron metabolic parameters with platelet counts in blood donors. METHODS: A total of 400 blood donors who met requirements of apheresis platelet donation were collected, and their hematological parameters were analyzed. The donors were divided into low ferritin group and normal group, the differences of hematological parameters between the two groups were compared, and the correlation of iron metabolic parameters and routine hematology parameters with platelet counts were analyzed. RESULTS: Whether male or female, low ferritin group had higher platelet counts than normal group (P < 0.01). Among the iron metabolic parameters, the platelet counts was negatively correlated with serum ferritin (SF), serum iron (SI), and transferrin saturation (TSAT) (r =-0.162, r =-0.153, r =-0.256), and positively correlated with total iron binding capacity (TIBC) and unsaturated iron binding capacity (UIBC) (r =0.219, r =0.294) in female blood donors. Platelet counts was also negatively correlated with SF, SI and TSAT (r =-0.188, r =-0.148, r =-0.224) and positively correlated with UIBC (r =0.220) in male blood donors. Among the routine hematology parameters, platelet counts was negatively correlated with mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and reticulocyte hemoglobin equivalent (Ret-He) in female blood donors (r =-0.236, r =-0.267, r =-0.213, r =-0.284). Platelet counts was also negatively correlated with MCH, MCHC and Ret-He in male blood donors (r =-0.184, r =-0.221, r =-0.209). CONCLUSION: In blood donors with low C-reactive protein level, the lower the iron store capacity, the lower the iron utilization, and the platelet counts tends to rise.

Enzyme stoichiometry evidence revealed that five years nitrogen addition exacerbated the carbon and phosphorus limitation of soil microorganisms in a Phyllostachys pubescens forest.

The activity and stoichiometry of soil extracellular enzyme can provide a good indication for changes in soil nutrient availability and microbial demands for nutrients. However, it remains unclear how would nitrogen (N) deposition affect nutrient limitation of microbes in subtropical forest soils. We conducted a 5 years N addition experiment in a subtropical Phyllostachys pubescens forest. The soil nutrients and enzyme activities associated with carbon (C), N, and phosphorus (P) cycles were measured. We also examined the nutrient distribution of microorganisms using enzyme stoichiometry and vector analysis. The results showed that N addition significantly decreased the contents of soil soluble organic C and available P and increased that of available N. Furthermore, N addition significantly decreased ß-N-acetyl-glucosaminidase (NAG) activity and NAG/ microbial biomass carbon (MBC), and increased acid phosphatase (ACP) and ACP/MBC. The low and moderate N addition levels significantly increased enzyme C/P, vector length, and vector angle, but significantly decreased enzyme N/P. Results of redundancy analysis showed that the change in soil enzyme activity and enzymatic stoichiometry were mainly driven by soil available P content under N addition. In summary, N addition altered the microbial nutrient acquisition strategy, which increased nutrient allocation to P-acquiring enzyme production but reduced that to N-acquiring enzyme production. Moreover, N addition exacerbated the C and P limitation of soil microorganisms. Appropriate amount of P fertilizer could be applied to improve soil fertility of subtropical P. pubescens forest in the future.

Tannins from senescent Rhizophora mangle mangrove leaves have a distinctive effect on prokaryotic and eukaryotic communities in a Distichlis spicata salt marsh soil.

Due to climate warming, tannin-rich Rhizophora mangle migrates into tannin-poor salt marshes, where the tannins interfere with the biogeochemistry in the soil. Changes in biogeochemistry are likely associated with changes in microbial communities. This was studied in microcosms filled with salt marsh soil and amended with leaf powder, crude condensed tannins, purified condensed tannins (PCT), all from senescent R. mangle leaves, or with tannic acid. Size and composition of the microbial communities were determined by denaturing gradient gel electrophoresis, high-throughput sequencing and real-time PCR based on the 16S and 18S rRNA genes. Compared with the control, the 16S rRNA gene abundance was lowered by PCT, while the 18S rRNA gene abundance was enhanced by all treatments. The treatments also affected the composition of the 16S rRNA and 18S rRNA gene assemblies, but the effects on the 18S rRNA gene were greater. The composition of the 18S rRNA gene, but not of the 16S rRNA gene, was significantly correlated with the mineralization of carbon, nitrogen and phosphorus. Distinctive microbial groups emerged during the different treatments. This study revealed that migration of mangroves may affect both the prokaryotic and the eukaryotic communities in salt marsh soils, but that the effects on the eukaryotes will likely be greater.

[Responses of soil phosphorus fractions and microorganisms to nitrogen application in a subtropical Phyllostachys pubescen forest]. / 亚热带毛竹林土壤磷组分和微生物对施氮的响应.

Phosphorus (P) is an important nutrient for plant and microbial growth. Soil P availabi-lity is poor in subtropical areas. Long-term heavy nitrogen (N) deposition might further reduce P availability. The experiment was performed in a Phyllostachys pubescens forest in Daiyun Mountain. The effects of N application on soil basic physical and chemical properties, soil P fractions, microbial biomass, and acid phosphomonoesterase activity were analyzed after three years of N application. The results showed that N application significantly increased NO3--N content and thus soil N availability, while it significantly reduced the percentage of decomposable organic P to total P, with the ratio of carbon (C) to organic P being over 200. The soil microbial biomass C, microbial biomass P, acid phosphomonoesterase, and the ratio of microbial biomass N to microbial biomass P and microbial biomass C to microbial biomass P were increased as the N application rate increased. There was a significant negative correlation between the percentage of decomposable organic P to total P and microbial biomass P. Consequently, N application enhanced soil P limitation and increased microbial P demand.

A Selective TRPC3 Inhibitor Pyr3 Attenuates Myocardial Ischemia/Reperfusion Injury in Mice.

An emerging body of evidence indicates that transient receptor potential TRP channels act as important mediators for a wide variety of physiological functions and are potential targets for drug discovery. Our previous study has identified transient receptor potential channel 3 (TRPC3) and TRPC6 as cation channels through which most of the damaging calcium enters, aggravates pathological changes in vivo and increases ischemia/reperfusion (I/R) injury in mice. This study aimed to verify the effects of TRPC3 inhibitor Pyr3 on myocardial I/R injury in mice. C57BL/6J wild-type male mice (8 to 12 weeks old) were anesthetized with 3.3% chloral hydrate. A murine I (30 min)/R (24 h) injury model was established by temporary occlusion of the left anterior descending (LAD) coronary artery. Pyr3 was administered at concentrations of 0, 2.5, 5, or 10 mg/kg via the right jugular vein 5 min before reperfusion. We observed that the selective TRPC3 inhibitor, 10 mg/kg Pyr3, significantly decreased the infarct size of left ventricle, and reduced the myocardial cell apoptosis rate and inflammatory response in mice. In a conclusion, TRPC3 can function as a candidate target for I/R injury prevention, and Pyr3 may directly bind to TRPC3 channel protein, inhibit TRPC3 channel activity, and improve TRPC3-related myocardial I/R injury. Pyr3 may be used for clarification of TRPC3 functions and for treatments of TRPC3-mediated diseases.

Nuclear FAM289-Galectin-1 interaction controls FAM289-mediated tumor promotion in malignant glioma.

BACKGROUND: FAM92A1-289(abbreviated FAM289) is recognized as one of the newly-discovered putative oncogenes. However, its role and molecular mechanisms in promoting cancer progression has not yet been elucidated. This study was performed to reveal its oncogenic functions and molecular mechanisms in human glioblastoma multiforme (GBM) cell models with knockdown or overexpression of FAM289 in vitro and in vivo. METHODS: To elucidate the molecular mechanisms underlying FAM289-mediated tumor progression, the protein-protein interaction between FAM289 and Galectin-1 was verified by co-immunoprecipitation, followed by an analysis of the expression and activity of Galectin-1-associated signaling molecules. Knockdown and overexpression of FAM289 in glioma cells were applied for investigating the effects of FAM289 on cell growth, migration and invasion. The determination of FAM289 expression was performed in specimens from various stages of human gliomas. RESULTS: FAM289-galectin-1 interaction and concomitant activation of the extracellular signal-regulated kinase (ERK) pathway participated in FAM289-mediated tumor-promoting function. Since the expression of DNA methyl transferase 1 (DNMT1) and DNA methyl transferase 3B (DNMT3B) was regulated by FAM289 in U251 and U87-MG glioma cells, Galectin-1 interaction with FAM289 may promote FAM289 protein into the cell nucleus and activate the ERK pathway, thereby upregulating DNMTs expression. Drug resistance tests indicated that FAM289-mediated TMZ resistance was through stem-like property acquisition by activating the ERK pathway. The correlation between FAM289, Galectin-1 expression and the clinical stage of gliomas was also verified in tissue samples from glioblastoma patients. CONCLUSIONS: Our results suggest that high expression of FAM289 in GBM tissues correlated with poor prognosis. FAM289 contributes to tumor progression in malignant glioma by interacting with Galectin-1 thereby promoting FAM289 protein translocation into the cell nucleus. FAM289 in the nucleus activated the ERK pathway, up regulated DNMTs expression and induced stem-like property gene expression which affects drug resistance of glioma cells to TMZ. This study provided functional evidence for FAM289 to be developed as a therapeutic target for cancer treatment.

[Effects of nitrogen deposition on the concentration and spectral characteristics of dissolved organic matter in soil in Moso bamboo plantations.] / æ°®æ²‰é™å¯¹æ¯›ç«¹æž—åœŸå£¤å¯æº¶æ€§æœ‰æœºè´¨æ•°é‡ä¸Žå ‰è°±å­¦ç‰¹å¾çš„å½±å“.

The subtropical zone in China is one of the regions most affected by nitrogen deposition. Soil dissolved organic matter (DOM) is considered to be an important indicator of soil organic matter. Nitrogen deposition may alter the quality and quantity of soil DOM by changing soil microbial activity. In this study, we explored the effects of nitrogen addition on soil DOM content, its spectral characteristics and microbial extraceller enzyme activity in the Moso bamboo plantations by setting control (CT), low-nitrogen (LN), and high-nitrogen (HN) addition levels for three-year nitrogen addition. The results showed that there was no significant change in soil pH, dissolved organic carbon, dissolved organic nitrogen, and aroma index following nitrogen addition, while the humification index increased significantly, microbial enzyme activities increased first and then decreased with the increases of nitrogen addition. Fourier transform infrared spectroscopy results showed that soil DOM had similar absorption peaks in seven regions, and that the absorption peaks of 1000 to 1260 cm-1 were the strongest, indicating an enhanced amount of polysaccharides, alcohols, carboxyl acids, and esters after nitrogen addition. The results of three-dimensional fluorescence spectroscopy showed that soil DOM structure significantly changed following nitrogen addition, with a decrease in low-molecular substances such as protein-like substances and microbial metabolites and a significant increase in high-molecular substances such as humus-like substances. In general, nitrogen addition made soil nitrogen compatible with microbial requirements. Microorganisms decompose substances that were easily degraded in DOM. The structure of soil DOM was more complex after nitrogen addition. Therefore, short-term nitrogen deposition might be conducive to preserving soil fertility.

[Nutrient and metabolic responses of the leaves of Cunninghamia lanceolata seedlings to warming and reduced precipitation in different season]. / ä¸åŒå­£èŠ‚æ‰æœ¨å¹¼è‹—å¶ç‰‡å »åˆ†å’Œä»£è°¢ç»„åˆ†å¯¹å¢žæ¸©å’Œå‡å°‘é™æ°´çš„å“åº”.

We examined the effects of warming (+5 ℃) and reduced natural precipitation (-50%) on nutrient status and physiological indices of Cunninghamia lanceolata seedlings during winter and summer in subtropical China. The results showed that seasonal changes in temperature and precipitation caused the seasonal differences in plant nutrient contents and metabolites levels. Contents of carbon, nitrogen, phosphorus, and potassium in leaves in winter were significantly higher than those in summer. In summer, reduced precipitation and warming had no significant effects on antioxidant enzyme activities in C. lanceolata leaves. In winter, superoxide dismutase and peroxidase activities in the leaves significantly decreased with reduced precipitation by 20.7% and 17.8%. Additionally, in winter, warming treatment significantly increased non-enzymatic ascorbic acid content by 132.5%. Carbon content decreased, whereas proline accumulation and nitrogen content increased under stress induced by combined warming and reduced precipitation in winter. However, carbon content increased by 3.3% under the treatment of simultaneous warming and reduced precipitation in summer. In addition, combined warming and reduced precipitation had no significant effects on the antioxidant system irrespective of the season. In conclusion, the adaptation mechanism of C. lanceolata to warming in summer might be different from that in winter. The changes in nutrient contents in C. lanceolata leaves were more sensitive to stress induced by combined warming and reduced precipitation. Nutrient demand and supply and seasonal changes in plant responses under climate change scenarios should be considered for better managing forest plantations and improving plant productivity.

[Effects of precipitation reduction on the composition and stability of soil organic matter in a young Cunninghamia lanceolata plantation.] / 减少降雨对杉木幼林土壤有机质组分及稳定性的影响.

It is hard to predict the response of soil organic matter (SOM) to global climate change due to its heterogenous chemical structure. With the development of molecular techniques to identify the structure, sources and stages of SOM degradation, long-standing questions regarding the composition and stability of SOM might be resolved. To investigate the effects of changes in precipitation patterns on the stability of SOM, we analyzed the specific compositions and extent of degradation of SOM using biomarkers, in a young Cunninghamia lanceolata plantation after reducing 50% of precipitation (P) for two years. The results showed that precipitation reduction (P-treatment) significantly reduced the levels of free lipids. Relative to control (CT), P-treatment decreased short-chain n-alkanoic acids (C16-18) and terpenoids and steroids by 62.8% and 19.1%, respectively. However, P-treatment did not significantly change the concentrations of other aliphatic compounds. Although there was no observable difference in the total lignin content between treatments, P-treatment significantly reduced the acid to aldehyde ratios for syringyl [(Ad/Al)s] and vanillyl [(Ad/Al)v]. Thus, the labile compositions of SOM were accelerated to decomposition under rainfall pattern change. Although the recalcitrant compositions (lignin) were relatively stable, their long-term stability should be further monitored.

Biatractylolide Modulates PI3K-Akt-GSK3ß-Dependent Pathways to Protect against Glutamate-Induced Cell Damage in PC12 and SH-SY5Y Cells.

Biatractylolide, isolated from the ethyl acetate extract of Atractylodes macrocephala, has shown various pharmacological activities such as antitumor and antioxidant activities. In this work, we aim to study the protective effect of biatractylolide on glutamate-induced rat adrenal pheochromocytoma cell (PC12) and human bone marrow neuroblastoma cell line (SH-SY5Y) injury and preliminarily explore its mechanism. The results showed that glutamate was cytotoxic with an inhibitory concentration 50% (IC50) of 8.5 mM in PC12 and 10 mM in SH-SY5Y cells. In this work, the preincubation with biatractylolide (10, 15, and 20 µM) observably improved cell viability, inhibited the apoptosis of cells induced by glutamate, and reduced the activity of LDH. AO staining revealed that apoptosis of cells was decreased. Additionally, the results of western blotting manifested that pretreatment with biatractylolide could downregulate GSK3ß protein expression and upregulate p-Akt protein expression, thereby protecting PC12 and SH-SY5Y cells from injury. All these findings indicate that biatractylolide has a neuroprotective effect on glutamate-induced injury in PC12 and SH-SY5Y cells through a mechanism of the PI3K-Akt-GSK3ß-dependent pathways.

Functional investigation on aromatase in endometrial hyperplasia in polycystic ovary syndrome cases.

OBJECTIVE: To explore the possible significance of aromatase P450 in endometrial hyperplasia with a background of polycystic ovary syndrome (PCOS). METHODS: Immunohistochemistry was used to determine the expression of aromatase P450 in endometrium of PCOS patients. Semiquantitative analysis of aromatase P450 expression of mRNA and protein level wasalso carried out by real-time quantitative RT-PCR method. After endometrial cells were stimulated by testosterone and letrozole in vitro, the estradiol (E2) level was determined, and the expression of cell aromatase P450 mRNA was assessed. RESULTS: The aromatase P450 mRNA level was increased in endometria of PCOS patients. When endometrial cells were cultured with 10-6 M testosterone, the E2 level in the culture medium increased. An inhibitory effect on E2 generation and expression of aromatase P450 mRNA was observed when the endometrial cells were treated with 10(-5) M letrozole. CONCLUSIONS: There is an increased expression of aromatase P450 in PCOS patient endometrium. Androgen stimulation could enhance the synthesis of aromatase P450 mRNA and the production of E2 in endometrial cells in vitro while letrozole could do the reverse.

Combination of serum inhibin B and follicle-stimulating hormone levels can not improve the diagnostic accuracy on testicular sperm extraction outcomes in Chinese non-obstructive azoospermic men.

BACKGROUND: It is still controversial whether the serum inhibin B level is a superior predictor of the presence of sperm in testicular sperm extraction (TESE) in azoospermic men compared with serum follicle-stimulating hormone (FSH). In this study, we evaluated the diagnostic accuracy of serum inhibin B levels as a predictor of the outcome of TESE in Chinese non-obstructive azoospermic men and compared it with the traditional marker serum FSH and testicular volumes. METHODS: Basal values of serum hormone levels, testicular volumes and histological evaluation of 305 Chinese non-obstructive azoospermic men were analyzed. The level of inhibin B was measured using a three-step enzyme-linked immunoassay before sperm extraction, and the diagnostic accuracy of prediction of the outcome of TESE was compared for different markers by the receiver operating characteristics (ROC) curve analysis. RESULTS: Testicular sperm was successfully retrieved in 137 of 305 patients (44.9%). The serum level of inhibin B, the FSH and the testicular volume were significantly different between the successful TESE group and the unsuccessful group. According to the ROC curve analysis, for inhibin B, the cut-off value for discriminating between successful and failed TESE was 28.39 pg/ml (sensitivity 83.5%, specificity 79.1%). For FSH, the best cut-off value for discriminating was 11.05 pg/ml (sensitivity 83.5%, specificity 74.5%). The area under the ROC curve of serum inhibin B was similar to that of FSH. Combining the serum inhibin B with FSH levels did not improve the predictive value for successful TESE. CONCLUSIONS: Serum inhibin B and FSH levels are correlated with spermatogenesis. However, inhibin B is not superior to FSH in predicting the presence of sperm in TESE. And the combination of them does not improve the diagnostic accuracy on TESE outcome.

Activation of epidermal growth factor receptor mediates reperfusion arrhythmias in anaesthetized rats.

AIMS: Epidermal growth factor receptor (EGFR) plays a critical role in the development and function of the heart. Previous studies have demonstrated that EGFR is involved in regulating electrical excitability of the heart. The present study was designed to investigate whether EGFR activation would mediate cardiac arrhythmias induced by reperfusion in anaesthetized rats. METHODS AND RESULTS: Reperfusion arrhythmias were induced by 10 min ligation of the left anterior descending coronary artery, followed by a 30 min reperfusion in anaesthetized rats. The incidence and severity of cardiac arrhythmias were significantly reduced by pre-treatment with the EGFR kinase inhibitor AG556. The phosphorylation level of myocardial EGFR was increased during ischaemia and at early reperfusion. Intramyocardial transfection of EGFR siRNA reduced EGFR mRNA and protein, and decreased the incidence of ventricular fibrillation induced by reperfusion. Interestingly, tyrosine phosphorylation levels of cardiac Na(+) channels (I(Na)) and L-type Ca(2+) channels (I(Ca,L)) were significantly increased at time points corresponding to the alteration of EGFR phosphorylation levels during reperfusion. AG556 pre-treatment countered the increased tyrosine phosphorylation level of Na(+) and L-type Ca(2+) channels induced by reperfusion. Patch-clamp studies proved that AG556 could inhibit I(Na) and I(Ca,L) in rat ventricular myocytes. No significant alteration was observed in tyrosine phosphorylation levels of cardiac Kv4.2 and Kir2.1 channels during reperfusion. CONCLUSION: These results demonstrate for the first time that EGFR plays an important role in the genesis of arrhythmias induced by reperfusion, which is likely mediated at least in part by enhancing tyrosine phosphorylation of cardiac Na(+) and L-type Ca(2+) channels.

Effect of curettage and copper wire on rabbit endometrium: a novel rabbit model of endometrial mechanical injury.

BACKGROUND: It remains almost a helpless situation for the recurrent implantation failure and pregnancy loss caused by endometrial injury at present. The purpose of this study was to develop a rabbit model of endometrial mechanical injury that could provide a research platform for this difficult clinical predicament. METHODS: Three experiments were conducted. Experiment 1: Curettages in both uterus horns and copper wire inserting after curettage (double-injury) in one horn. The histological changes were monitored at 0, 24, 48, 72 hours, as well as in 1 and 2 weeks after operation. Experiment 2: Direct copper wire inserting in one horn and double-injury in other horn. The wires in both horns were removed after 2 weeks. The histological changes were recorded at 0, 1 and 2 weeks after wire removal. Experiment 3: Double-injury procedure in one horn was performed and wire was removed after 2 weeks; another horn was remained normal to serve as control. Histological changes were recorded, tissue areas were measured, and proliferation indices (PIs, %) were calculated at 1, 2, 4 and 8 weeks after wire removal, respectively. RESULTS: The experiments revealed that the injured endometrium by simple curettage or copper wire could be fully repaired. While the endometrial regeneration was severely impaired by double-injury, both areas of endometrium and uterine cavity decreased (P < 0.05); both PIs of glandular epithelial and stromal cells increased and reached maximum at 4 weeks (P < 0.05), but returned by 8 weeks. CONCLUSION: This study demonstrated that a rabbit model of endometrial injury could be effectively established through a double-injury procedure of curettage and copper wire with comparable clinical index.

Alteration of ERß gene RsaI polymorphism may contribute to reduced fertilization rate and embryonic developmental competence.

This paper aims to determine the possible role of estrogen receptor-ß (ERß) gene RsaI polymorphism on sperm fertility and early embryonic development in humans. Three groups of Chinese men were recruited: in vitro fertilization (IVF) group, including 374 couples who underwent conventional IVF; intracytoplasmic sperm injection (ICSI) group, including 294 couples who underwent an ICSI procedure using ejaculated sperm; and azoospermic group, consisting of 197 couples who underwent ICSI using either testis or epididymis sperm. RsaI polymorphism in the ERß gene was detected by polymerase chain reaction (PCR)-restriction fragment length polymorphism technique; fertilization and high-quality embryo rates were evaluated for each group. In each group, no significant differences were found in the overall rates of fertilization and high-quality embryos among GG, AG and AA genotypes. However, the proportion of cycles possessing a satisfactory high-quality embryo rate with the AA genotype was significantly lower than that in the wild-type GG genotype from each group. These results demonstrated that sperm possessing the ERß RsaI A genotype may have reduced fertilization ability and decreased early embryonic developmental potential, which could directly or indirectly contribute to the low fertilization rate and early embryonic developmental arrest in some cases.

[Study of HBV-DNA in ovarian granulosa cells of hepatitis B carriers].

OBJECTIVE: To study the infection status of hepatitis B virus in ovarian granulosa cells of female hepatitis B carriers. METHODS: The follicular fluid of 6 female carriers of hepatitis B (HBsAg+, HBeAg+, HBcAb+) was used to extract granule cells and prepare smears. The samples were detected via in situ hybridization by digoxin-labeled HBV-DNA probe. The color reaction system of alkaline phosphates was used. And the positive hybridization signal was purple blue. RESULTS: Three granulosa cell samples, located at the nuclei of granulosa cells, were HBV-DNA positive. CONCLUSION: HBV DNA exists in the granulosa cells of hepatitis B patients. Thus HBV infection may be vertically transmitted to the offspring via the infected reproductive cells.