Effects of changing PKC activity on proliferation and telomerase expression in human hepatocellular carcinoma cells

AIM: To investigate whether protein kinase C (PKC) is involved in the proliferation and the telomerase expression in human hepatocellular carcinoma cells. METHODS: Human hepatocellular carcinoma cells (BEL-7402) were treated with exogenous phorbol-12-myristate-13-acetate (PMA, PKC activator) and staurosporine (SP, PKC inhibitor) for 48 hours. The techniques of cell culture and the telomeric repeat amplification protocol silver staining in combination with computer image scanning system in vitro were used to observe the variations of the growth and the telomerase expression. RESULTS: The proliferative potential of BEL-7402 cells was decreased by the action of PMA as well as SP, and the telomerase expression was also inhibited by PMA and SP. CONCLUSION: Our findings suggest that the proliferation of human hepatocellular carcinoma cells and the telomerase expression may be related to PKC.     

Effect of YIGU capsule on IGF-I mRNA and protein expression in rat osteoblasts in vitro

AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P<0.01); (2) The relative IGF-I mRNA levels and IGF-I protein expression were higher than those in control group after 48, 72 and 96 h, respectively (P<0.01 or P<0.05). CONCLUSIONS: It was suggested that YIGU capsule drug-containing serum promoted proliferation, IGF-I mRNA and protein expression. These results may be parts of the mechanisms of YIGU capsule to prevent and treat osteoporosis.     

Effects of fosinopril,captopril and valsartan on the expressionof tissue factor in human monocytes

AIM: To investigate the effects of angiotensin-converting enzyme inhibitors (ACEI), fosinopril, captopril and angiotensin II AT₁ antagonists, valsartan on tissue factor (TF) expression on monocytes induced by lipopolysaccharide (LPS). METHODS: Mononuclear leukocytes from normal delivered female umbilical veins were incubated with bacterial LPS in presence or absence of different ACE inhibitors .At the end of incubation, the cells were disrupted by 3 freeze-thaw cycles. TF procoagulant activity was assessed by a one-stage clotting assay. RT-PCR was used to check TF mRNA expression, and GAPDH mRNA was used for parallel assay. RESULTS and CONCLUSION: The results showed that increased expression of TF mRNA induced by LPS was inhibited by fosinopril, captopril and valsartan, respectively, and the procoagualant activity of monocytes was also reduced.     

Effects of β-ME and RA on expression of GFAP in mesenchymal cells derived from mouse fetal liver

AIM: To investigate the effects of β-mercaptoethanol (β-ME) and all-trans rentinal acid (RA) on glial fibrillary acidic protein (GFAP) expression in mesenchymal cells derived from mouse fetal liver in vitro. METHODS: Cells suspension from 14.5-days-old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20％ FCS and mesenchymal cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by β-ME and RA. The characteristics of treated cells were assayed by immunocytochemistry staining at 5 hours and 5 days after induction. β-actin as an internal control, GFAP gene expression of mesenchyal cells was detected with semi-quantitative RT-PCR. RESULTS: After being inducted by β-ME and RA, 80％ approximately of the cells exhibited typical neural morphology and about 85％ expressed GFAP phenotype. Semi-quantitative RT-PCR showed that mRNA expression of GFAP increased in treated cells versus untreated cells (P<0.01). CONCLUSION: GFAP expression in mesenchymal cells derived from mouse fetal liver in vitro increases after being treated with β-ME and RA.     

羊、鸡抗鼠脂肪细胞膜抗血清对大鼠体脂与生长性能的影响

应用提取的鼠脂肪细胞膜分别免疫羊和鸡 ,所产生的抗血清用于 Wistar大鼠被动免疫。实验 1: 组腹腔注射羊正常血清 , 组腹腔注射羊抗鼠脂肪细胞膜抗血清 ,剂量均为 1m L /只 ,连续注射 4 d。结果表明 ,羊抗鼠脂肪细胞膜抗血清免疫促进了大鼠体增重 ,降低了体脂沉积 ,与对照组相比 ,7周末体重增加 6 .35 % (P<0 .0 5 ) ,饲料摄入增加6 .85 % (P<0 .0 1) ,料重比 (F/G)提高 4 5 .0 0 % (P<0 .0 5 ) ;肾周、附睾、网膜脂肪垫重量分别降低 2 3.92 % (P<0 .0 5 )、34.4 5 % (P<0 .0 5 )、0 .98% ,脂肪总量降低 2 0 .92 %。实验 2 :1组腹腔注射鸡正常血清 ,2组腹腔注射鸡抗鼠脂肪细胞膜抗血清 ,剂量均为 1m L/只 ,连续注射 4 d。结果表明 ,鸡抗鼠脂肪细胞膜抗血清免疫对大鼠的生长发育产生了不利影响 ,7周末 ,免疫大鼠平均体重较对照组减少 4 0 g(P<0 .0 5 ) ,饲料摄入显著降低 (P<0 .0 1) ;对体脂的沉积和血液中 TG和 FFA的影响没有规律 ,且无统计学意义     

Cloning and characterization of a bovine genomic fragment homologous to epidermal growth factor genes

Epidermal growth factor (EGF) is a potent mitogen for a variety of cell types. The 53-amino acid mature EGF protein is encoded by sequences in exons 20 and 21 of a gene spanning over 110 kb. In this study, we report the cloning and characterization of 7.5 kb of bovine genomic sequence homologous to exon 19 through 21 from EGF genes from other mammalian species. The cloned gene fragment had an unusual sequence composition in the form of an in-frame TGA codon in the coding sequence. The sequence was expressed at low levels in kidney tissue and the corresponding cDNA contained the TGA codon. The level of similarity between the bovine exonic sequence and the human, porcine, murine, feline, and canine corresponding sequences varied from 64% to 73%; however, when only sequences encoding the mature EGF protein were compared, the level of similarity between the bovine sequence and the sequence from these species was 59% to 66%. The sequence similarity of the deduced mature protein was lower (34% to 39%) than the sequence similarity of the deduced propeptide. Although the cloned sequences could originate from a bovine EGF pseudogene, the possibility exists that they originate from the functional EGF gene. An as yet unidentified mechanism to by-pass the stop codon would allow the synthesis of a functional EGF protein. Alternatively, the cloned sequence could originate from an EGF-like gene.     

Determination of plasma lysosomal enzyme activity and its relationship to severity of injury following blunt vehicular trauma in dogs

Objective: To determine plasma β‐d ‐glucuronidase (βG) activity in the first 4 hours following injury in dogs struck by a motor vehicle, and to evaluate whether the degree of enzyme activity is correlated with the severity of injury. Design: A prospective clinical study. Setting: Veterinary Medical Teaching Hospital. Animals: Thirteen client‐owned dogs that were presented to the Veterinary Hospital of the University of Pennsylvania between June and August 1999 for blunt vehicular trauma. Ten healthy student and staff‐owned dogs served as controls. Interventions: None. Measurements: Plasma was analyzed for βG enzyme activity at the time of presentation (n=13), and 1 and 4 hours (n=7) following presentation to the Emergency Service for blunt vehicular trauma. The results were compared with enzyme activity from healthy controls evaluated serially over 4 hours. Fluorometric analysis using 96‐well microtiter plates was used to perform the enzyme assays. The relationships between presentation (n=13) and 4 hours (n=7) of enzyme activity and 3 indices of metabolic and physical disturbance (serum pH, serum lactate and Animal Trauma Triage (ATT) score) at the time of presentation were also investigated. Main results: Of the 13 dogs, 7 fulfilled the inclusion criteria for comparison of enzyme activity of the trauma over time. A statistically significant difference in βG activity was found in the trauma group (mean 75.6±10.4 U) at 4 hours following presentation compared with controls (mean 48.0±6.4 U). This difference was suggested by 1 hour following presentation (trauma group, mean 70.4±10.9 U; control group, mean 49.8±5.5 U), although it did not reach statistical significance. Thirteen dogs fulfilled the inclusion criteria for comparison of only presentation enzyme activity with trauma severity score, serum lactate, and serum pH. No statistically significant relationship was found between the βd ‐glucuronidase activity and the presenting ATT score, serum lactate concentration, or serum pH at either presentation or 4 hours, although the power of these analyses was low. Conclusions: These results demonstrate that the activity of βG, a lysosomal enzyme, increases significantly in the systemic circulation in dogs 4 hours following blunt trauma. Additional research to include more severely injured dogs, a larger number of dogs, and to follow the course of injury for a longer period of time would be beneficial to further characterize βG activity following blunt trauma.     

Association of hyponatremia and hyperglycemia with outcome in dogs with congestive heart failure

Objective: To evaluate plasma sodium and glucose concentrations in dogs with congestive heart failure (CHF) prior to treatment and evaluate the differences between survivors and non‐survivors. Design: Retrospective study. Animals: Fifty‐nine dogs with CHF prior to receiving cardiac medication. Interventions: None. Measurements and main results: The mean plasma sodium concentration in dogs with CHF was below the reference range (144–156 mmol/L) and significantly lower (P=0.009) in non‐survivors (141±6 mmol/L) compared with survivors (147±4 mmol/L). The mean plasma glucose concentration was above the reference range (76–117 mg/dL) and significantly higher (P=0.004) in non‐survivors (128±52 mg/dL) compared with survivors (100±13 mg/dL). Forty‐four percent of non‐survivors had concurrent low plasma sodium and high plasma glucose concentrations, whereas no survivors had both abnormalities (P<0.0001). Conclusions: Lower plasma sodium and higher plasma glucose are associated with a worse outcome in dogs with CHF.     

A computer program for calculation of doses and prices of injectable medications based on body weight or body surface area

A computer program (CalcAnesth) was developed with Visual Basic for the purpose of calculating the doses and prices of injectable medications on the basis of body weight or body surface area. The drug names, concentrations, and prices are loaded from a drug database. This database is a simple text file, that the user can easily create or modify. The animal names and body weights can be loaded from a similar database. After typing the dose and the units into the user interface, the results will be automatically displayed. The program is able to open and save anesthetic protocols, and export or print the results. This CalcAnesth program can be useful in clinical veterinary anesthesiology and research. The rationale for dosing on the basis of body surface area is also discussed in this article.     

鸭疫里氏杆菌形态特征的电镜观察

自鸭传染性浆膜炎的病死鸭中分离鉴定的鸭疫里氏杆菌(RA)Ⅰ、Ⅱ型两个菌株，用负染法和超薄切片法制备样品，于透射电镜下进行观察。结果观察到约占1／3的RA具有特殊的形态结构：在细菌的顶端或侧面有1～2个与菌体相连的芽状赘生物，有的也能从菌体上脱落下来。它在超薄切片中的大小约为菌体的1／3～1／5，能观察到其内部也具有类似菌体内部的结构。且具有类似菌体的“细胞壁”，类似于核体部分的结构位于“细胞壁”的一侧。而用相同的方法制作的鸭大肠杆菌(E．cozi)对照样品，却见不到这种特殊的形态结构。说明这种形态特征可能是RA特有的。