Comparison of glomerular filtration rate between greyhounds and non-Greyhound dogs

Greyhounds have significantly higher serum creatinine (SCr) concentration than do non-Greyhound dogs that may be attributable to differences in glomerular filtration rate (GFR). By means of plasma clearance of technetium Tc 99m diethylenetriaminepentaacetic acid, GFR was measured in 10 Greyhounds and 10 non-Greyhound dogs with normal findings of physical examination, CBC, serum biochemical analysis, and urinalysis. Dogs were fed the same diet for a minimum of 6 weeks before GFR data collection. Greyhounds had significantly higher mean +/- SD GFR (3.0 +/- 0.1 vs 2.5 +/- 0.2 ml/min/ kg; P = .01) and SCr concentration (1.8 +/- 0.1 vs 1.5 +/- 0.1 mg/dL; P = .03) than did non-Greyhound dogs, but the serum urea nitrogen (SUN) concentration was not significantly different (18 +/- 1 vs 18 +/- 2 mg/dL; P = .8). Therefore, the higher SCr concentration in Greyhounds is not attributable to decreased GFR, and may be associated with the high muscle mass in the breed. Healthy Greyhounds have higher GFR than do non-Greyhound dogs.     

Comparison of ventral coccygeal arterial and jugular venous blood samples for pH, pCO2, HCO3, Be(ecf) and ctCO2 values in calves with pulmonary diseases

The aim of this study was to determine whether venous blood samples can be used as an alternative to arterial samples in calves with respiratory problems and healthy calves. Jugular vein and ventral coccygeal artery were used to compare blood gas values. Sampling of the jugular vein followed soon after sampling of the ventral coccygeal artery in healthy calves (group I) and calves with respiratory problems (group II). Mean values of arterial blood for pH, pCO2, HCO3act in healthy calves were 7.475 +/- 0.004, 4.84 +/- 0.2 kPa, 28.45 +/- 1.30 mmol/L compared with venous samples, 7.442 +/- 0.006, 6 +/- 0.3 kPa, 30.93 +/- 1.36 mmol/L, respectively. In group II, these parameters were 7.414 +/- 0.01, 5.93 +/- 0.3, 27.73 +/- 1.96 mmol/L for arterial blood and 7.398 +/- 0.008, 6.85 +/- 0.2 kPa, 29.77 +/- 1.91 mmol/L for venous blood, respectively. There were no statistically significant differences between arterial and venous pH, HCO3act, Be(ecf), ctCO2 values with the exception of pCO2 (P = 0.001) in group II. In group I, correlation (r2) between arterial and venous blood pH, pCO2, HCO3act were 84.5%, 87.5%, 95.7%, respectively compared with the same parameters in group II, 80.8%, 77.1%, 70.3%. In conclusion, venous blood gas values can predict arterial blood gas values of pH, pCO2 and HCO3ecf, Be(ecf) and ctCO2- for healthy calves but only pH values in calves with acute respiratory problems (r2 value>80%).     

Regulatory effects of Rock2 on cell cycle checkpoint Cdc25A

AIM: To investigate the regulatory effects of Rho-associated coiled-coil-containing protein kinase 2(Rock2) on the cell cycle checkpoint cell division cycle 25A(Cdc25A). METHODS: The protein expression levels of Rock2 and Cdc25A in 51 pairs of hepatocellular carcinoma and the adjacent tissues were detected by Western blotting. shRock2 plasmids were constructed, selected and stably transfected into hepatocellular carcinoma Huh-7 and HepG2 cells. The protein expression of Cdc25A in the cells was determined by Western blotting. Based on the Rock2 interfering sequences, we designed the primers and changed the 4 indicated bases via site-specific mutagenesis. The Rock2-mutant plasmid was verified by sequencing and was transfected into stable Rock2-knockdown cells. The protein expression of Cdc25A was detected by Western blotting, and the cell proliferation was measured by MTT assay. The protein levels of checkpoint kinase(Chk)1/Chk2 were also detected in stable Rock2-knockdown cells. The interaction between Rock2 and Cdc25A was measured by co-immunoprecipitation, and the co-localization of Rock2 and Cdc25A was detected by confocal laser scanning microscopy. RESULTS: Rock2 and Cdc25A were apparently up-regulated in hepatocellular carcinoma,with a significantly positive correlation. The protein expression of Cdc25A was significantly down-regulated in stable Rock2-knockdown cells. The expression of Chk1 and Chk2 was not changed following knockdown of Rock2. The co-immunoprecipitation results verified that Rock2 bound to Cdc25A. The results of confocal laser scanning microscopy showed that Rock2 and Cdc25A were co-localized in hepatocellular carcinoma cells. CONCLUSION: Rock2 positively regulates the cell cycle checkpoint Cdc25A, which is independent of Chk1/Chk2 and this may provide a new target gene for treatment of hepatocellular carcinoma.     

Effects of edible-medicinal fungus crude extracts on vascular endothelial cells with high glucose-induced injury

AIM: To investigate the effects of crude extracts of Cordyceps gunnii (CGE), Lepista lentinus (LLE), Cordyceps sinensis (CSE) and Lentinus striguellus (LSE) on the proliferation of high glucose-treated human umbilical vein endothelial cells (HUVECs). METHODS: The cultured HUVECs were divided into normal control group (treated with M199 culture medium alone), high glucose group (treated with M199 culture medium containing 33 mmol/L glucose) and 4 crude extracts of edible-medicinal fungi (CGE, LLE, CSE and LSE) intervention groups (treated with the crude extract of edible-medicinal fungus at concentrations of 12.5, 25, 50, 100 mg/L in high glucose M199 culture medium). The cell proliferation was evaluated by MTT assay. The cell cycle and ROS level were measured by flow cytometry. RESULTS: Compared with normal control group, the MTT absorbance value and the percentage of G₀/G₁ stage of the HUVECs in high glucose group were significantly decreased (P<0.05), while the percentage of S+G₂/M and ROS level were significantly increased (P<0.05). Compared with high glucose group, treatment with the crude extracts of Lepista lentinus and Lentinus striguellus decreased the cell absorbance value (P<0.05), and the inhibitory effect was enhanced in a dose-dependent manner. However, Cordyceps gunnii had no effect (P>0.05). The crude extracts of Cordyceps sinensis (12.5~50 mg/L) significantly enhanced the proliferative activity of HUVECs, decreased the percentage of G₀/G₁ stage of HUVECs, increased the percentage of S+G₂/M of HUVECs, and reduced the intercellular ROS level (P<0.05). CONCLUSION: Only Cordyceps sinensis crude extract effectively protects the HUVECs in high glucose-induced injury, which might be due to promoting more cells to enter to the cell cycle and down-regulating oxidative stress.     

Effects of adenovirus-mediated shRNA targeting PTEN on proliferation and apoptosis of activated hepatic stellate cells in vitro

AIM:To investigate the down-regulation of phosphatase and tensin homolog deleted on chromosome 10(PTEN) gene by adenovirus-mediated short hairpin RNA(shRNA) on proliferation and apoptosis of activated hepatic stellate cells(HSCs) in vitro and the related signaling transduction pathways. METHODS:The activated HSCs were cultured in vitro and transfected with recombinant adenovirus expressing shRNA targeting PTEN. The proliferation of HSCs was measured by MTT assay and the apoptosis was assessed by TUNEL and flow cytometry. Western blotting was used to detect the protein levels of PTEN, Bax, Bcl-2, Akt, p-Akt, ERK1/2 and p-ERK1/2 in HSCs, and real-time fluorescent quantitative PCR was applied to detect the mRNA expression of Akt and ERK1. RESULTS:The recombinant adenovirus expressing shRNA targeting PTEN was successfully transfected into activated HSCs in vitro, and significantly promoted the proliferation of HSCs in a time-dependent manner within a certain extent. The apoptotic rate of HSCs was significantly decreased 72 h after transfection(P<0.05). Meanwhile, reduced expression of Bax and elevated expression of Bcl-2 were induced 72 h after transfection(P<0.05). Furthermore, the expression of p-Akt and p-ERK1/2 were increased significantly(P<0.05), while no significant difference in the expression of Akt and ERK1 at mRNA and protein levels was observed(P>0.05). CONCLUSION:Down-regulation of PTEN by adenovirus-mediated shRNA dramatically promotes the proliferation of activated HSCs, and inhibits the apoptosis through Bcl-2/Bax pathway. In addition, the phosphorylation of Akt and ERK1/2 is increased, indicating that PI3K/Akt and ERK1/2 signal transduction pathways may play an important role in the regulation of proliferation and apoptosis of HSCs.     

红与黑西瓜新品种的选育和推广

红与黑是合肥丰乐种业股份有限公司选育的椭圆形黑皮红瓤优质西瓜品种。中熟品种,全生育期110d,果实发育期33d左右。植株长势稳健,易坐果且坐果整齐。主蔓第1朵雌花着生于7节左右,以后每隔6—7节再现1朵雌花。果形指数约1．3,大红瓤,中心可溶性固形物12．2％左右,口感好,品质佳。单果质量6-8kg,667m。产量3500kg以上。皮厚1．2cm,耐贮运。适宜全国露地西瓜种植栽培。     

Effects of human growth arrest-specific homeobox gene delivery on proliferation,migration and cell cycle distribution of vascular smooth muscle cells

AIM: To explore a new gene therapeutic strategy for vein graft restenosis by investigating the effects of adenovirus-mediated human growth arrest-specific homeobox (Ad5-hGax) gene delivery on the proliferation, migration and cell cycle distribution of serum-induced rabbit vascular smooth muscle cells (VSMCs). METHODS: The recombinant adenovirus vector containing hGax gene was constructed and transfected into rabbit VSMCs. The expression of hGax in VSMCs was detected by RT-PCR and immunofluorescent staining. Methyl thiazolyl tetrazolium (MTT) assay was used to assess the effect of hGax over-expression on serum-induced proliferation of VSMCs. Wound healing method was applied to examine the distance of serum-induce VSMCs migration. Flow cytometry was employed to analyze the cell cycle distribution. RESULTS: The recombinant adenovirus vector Ad5-hGax was successfully constructed. The results of RT-PCR and immunofluorescent staining revealed that the hGax -transfected cells contained a 174 bp specific fragment of hGax gene and target protein 48 h after transfection. The proliferation of serum-induced VSMCs was significantly inhibited by overexpression of hGax gene as compared with control group. The migration of serum-induced VSMCs was inhibited after hGax gene delivery. Flow cytometry showed that 72 h after serum induction, the cells in G₀/G₁ phase in Ad5-hGax group were significantly increased, whereas the cells in G₂/M+S phase were significantly decreased. CONCLUSION: Overexpression of hGax gene inhibits the proliferation and migration of serum-induced rabbit VSMCs, and arrests the cells in G₀/G₁ phase. It is likely that hGax gene is a potential target for the gene therapy of vein graft restenosis.     

Nerve growth factor promotes proliferation of hepatocytes in vitro via TrkANGFR signal pathway

AIM: To observe the expression and distribution of nerve growth factor (NGF), its high-affinity tyrosine kinase A receptor (TrkA^NGFR) and low-affinity p75 neurotrophin receptor (p75^NTR) in hepatocytes(L02 cells), and to investigate the effects of exogenous recombinant human NGF-β on L02 cells. METHODS: L02 cell line was used in the experiment. The expression and intracellular distribution of NGF, TrkA^NGFR and p75^NTR were detected by the methods of immunocytochemistry and fluorescent quantitative PCR. The proliferation of L02 cells after exposed to exogenous NGF-β, anti-NGF, anti-TrkA^NGFR and anti-p75^NTR was detected by XTT {2,3-bis(2-methoxy-4-nitro-5- sulfophenyl)-5- -2H-tetrazolium hydroxide} assay. The cell apoptosis and the change of cell cycle of L02 cells after exposed to exogenous NGF-β were determined by flow cytometry with Annexin V-FITC and PI staining. RESULTS: The expression of NGF was mainly localized in the cytoplasm and nuclei of L02 cells. The expression of TrkA^NGFR was highly and the expression of p75^NTR was weakly detected in the cell membrane and cytoplasm of L02 cells. Exogenous NGF-β promoted the expression of NGF and TrkA^NGFR in L02 cells. Low dose of exogenous NGF-β (12.5-200 μg/L) promoted L02 cell proliferation and inhibited the cell apoptosis by affecting the cell cycle in S-phase. High dose of NGF-β (>400 μg/L) did not promote L02 cell proliferation. Specific neutralizing antibodies of NGF and TrkA^NGFR decreased the NGF-β-induced proliferation of L02 cells. However, blocking the binding of NGF to p75^NTR by anti-p75^NTR did not affect the NGF-β-induced proliferation of L02 cells. CONCLUSION: L02 cells express NGF, TrkA^NGFR and p75^NTR. Exogenous recombinant human NGF-β at optimal dose promotes L02 cell proliferation in a dose-dependent manner possibly via NGF/TrkA^NGFR signal pathway.     

Lovastatin-induced G1 phase synchronization in human endometrial cancer JEC cells

AIM: To investigate the method of inducing G₁ phase synchronization in human endometrial cancer JEC Cells by lovastatin and the cell cycle progress of JEC cells after desynchronization. METHODS: The doubling time of JEC cells was detected by Cell Counting Kit-8 (CCK-8) assay. To determine the best lovastatin concentrations for G₁ synchronization, JEC cells were treated with lovastatin at concentrations of 10, 20, 30 and 40 μmol/L for 1× doubling time, and the cell cycle was detected using flow cytometry (FCM). To determine the best period of lovastatin treatment to achieve G₁ synchronization, JEC cells were treated with lovastatin at the best concentration for 0.5× to 2× doubling time, and the cell cycle was detected every 4 h using FCM. Furthermore, the cell cycle progress of JEC cells after desynchronization was also observed. RESULTS: The doubling time of JEC cells was almost 24 h. Treatment with lovastatin at the concentration of 20 μmol/L for 24 h achieved maximum G₁ arrest in JEC cells. Minimum G₁ phase and maximum S phase were observed after desynchronization for 16 h. CONCLUSION: Maximum G₁ synchronization of JEC cells is induced by lovastatin at the concentration of 20 μmol/L for 24 h. The JEC cells show minimum G₁ phase and maximum S phase after desynchronization for 16 h.