RESEARCH ON LACTIC DEHYDROGENASES (LDH) ISOENZYMES OF FOUR SPECIES IN GALLIFORMES

Low-ion-density discontinuous polyacrylamide electrophoresis (pH 8.0) of LDH from different tissuse of four species (Tetrastes bonasia,Chrysolophus pictus, Phasians colchicus, Gallus gallus domesticus) in Galliformes showed that this electrophoresis system was suitable for analysing avian LDH. Thermostabilty to heat and nonsusceptibility to urea inhibition of low-density urea of the four liver LDHs were in such order as LDH1> LDH2> LDH3> LDH4> LDH5. And it appeared. by comparing relative movable ratio of A and B subunits of LDHs, that difference exists between Phasianidae and Tetraonidae and in Phasianidae Phasianus was closer to Chrysolophus than to Gallus.

Effect of Purified Paper Wasp Ropalidia marginata Venom Toxin Enzyme Activity in Blood Serum, Liver, and Gastrocnemius Muscle of Albino Mice

In the present, investigation effects of sub-lethal dose of purified paper wasp Ropalidia marginata venom toxins were evaluated on important metabolic enzymes i.e. ALP ACP, GPT, GOT, LDH, and AchE enzyme activity in serum, liver, and gastrocnemius muscles of albino mice. Alkaline phosphatase was found to be increased up to 119.9% at the 6th hr of the toxin injection in comparison to control. This elevation may be due to cytolysis. Maximum increase i.e., 153.33% level of glutamate pyruvate transaminase (GPT) was found at 6 hrs of 40% of 24-h LD50 treatment while it was found to be 151.1% at 6 hrs of 24 hr 80% of LD50, venom injection. A significant elevation was observed in LDH activity in serum, liver, and muscles, while the activity of AchE was decreased in serum, liver, and gastrocnemius muscles of albino mice after injecting the sub-lethal dose of Ropalidia marginata venom. This increase in the activity of LDH produces liver damage, massive disintegration and necrosis of hepatic cells. This elevation in LDH level led to a significant increase in the glucose catabolism and elevated oxidative stress in muscle and liver cells. It also displays insufficient oxygen supply and consequently leads to cell death. In experimental animals, venom toxin treatment decreased AchE level, and animals showed muscular paralysis. When mice were treated with 40% and 80% of 24-h LD50 of purified venom caused a significant (p < 0.05) elevation in the level of ACP, GOT, GPT, and LDH while the reduction in ALP and AChE level. Present study will be useful in the development of prototypes for study of pharmacological and therapeutic effects of various venom toxins. For this purpose structure activity relationship of enzyme and venom toxin, its due interaction to various metabolic enzymes and receptors must be explored.

Effects of Purified Indian Cattle Tick Rhipicephalus microplus Saliva Toxins on Various Enzymes in Blood Serum, Liver and Muscles of Albino Mice

In the present investigation, in vivo effects of purified ticks’ saliva toxin were evaluated on the level of certain important cellular metabolic enzymes i.e. acid phosphatase (ACP), alkaline phosphatase (ALP), glutamate pyruvate transaminase, glutamate oxaloacetate transaminase and lactic dehydrogenase. For this purpose, sub-lethal doses, 40% and 80% of 24 h LD50 purified saliva toxins of Rhipicephalus microplus (Canestrini, 1888) were injected subcutaneously in the albino mice. In treated mice saliva toxins targeted membrane-bound enzymes i.e. serum acid phosphatase and alkaline phosphatase, its level was increased from 118.30% to 163.63% at the 6th hr in comparison to the control. Besides this, the levels of serum glutamate pyruvate transaminase (GPT) and glutamate oxaloacetate transaminase (GOT) and lactic dehydrogenase (LDH) also increased up to 161.11% (at 6th hr), 148.27 (at 8th hr) and 125.45% (at 6th hr) respectively in comparison to control. An increase in the level of LDH showed insufficient oxygen supply, massive disintegration of cells and leakage of the enzyme into the circulation. It clearly indicated the toxic effects of saliva toxins on the membrane of blood cells, hepatocytes and myocardial muscle cell functions in albino mice. On the other hand activity of acetyl cholinesterase was reduced by 65.51% at the 6th hr of the saliva toxin injection in comparison to the control. This inhibition of acetyl cholinesterase activity caused the accumulation of acetylcholine molecules at the synaptic junctions and led to prolonged activation of acetylcholine receptors. It caused permanent stimulation of nerves and muscle cells that may result in muscular paralysis and finally death of the animal.

Effect of adrenalectomy and hydrocortisone on ventral prostate of rats

Aim: To study the effects of adrenalectomy and hydrocortisone on the ventral prostate of SD rats. Methods: Inadrenalectomised (ADX) and ADX + hydrocortisone (1, 2, or 4 mg) treated rats, the prostatic histology and thecholesterol, protein, zinc, and copper levels and the enzymic profile (acid phosphatase, alkaline phosphatase, aryl sul-phatase, lactic dehydrogenase, and leucine aminopeptidase) in the prostatic tissue were determined; the serum hormon-al profile (testosterone, FSH and LH) was also assayed. Results; Adrenalectomy caused a progressive degenerationin prostatic structure that was not reversed by hydrocortisone treatment. The serum testosterone were significantly lowerin ADX than in sham operated rats and lower in ADX + hydrocortisone than in ADX-C rats (P < 0.01). The serumFSH and LH were below the detection limit of 1 mIU/mL. The enzymatic activity was higher in ADX than in sham op-erated rats and higher in ADX + hydrocortisone than in ADX-C rats (P<0.05-0.01). The prostatic zinc levels weresignificantly higher in sham operated than in ADX, and higher in ADX-C than in ADX + hydrocortisone rats (P < 0.05-0.01). The prostatic copper level was significantly lower in sham operated than in ADX, and lower in ADX-C thanin the ADX + hydrocortisone rats (P <0.01). Conclusion; In rats, adrenalectomy leads to pathological and func-tional changes of the prostate. Hydrocortisone treatment at the doses employed did not reverse these changes. (Asian JAndrol 2001 Dec; 3: 289 - 300)

Thick tongue coating: diagnostic markers for metastatic colorectal cancer?

Background:According to World Health Organization,colorectal cancer is the third most common cancer in the world.The prognosis assessment and condition judgment of the colorectal cancer remains a challenge clinically.Therefore,identification of diagnostic markers to evaluate the prognosis of colorectal cancer clinically should be urgently developed.We have observed that a lot of cancer patients had thick tongue clinically,but what is the relationship between tongue coating and the tumor?Methods:Seventy-four patients with colorectal carcinoma were collected through the outpatients of Tianjin Medical University Cancer Institute and Hospital from May 2010 to September 2011,in which there were 49 patients confirmed with recurrence or metastasis.All photos of patients’tongue were taken with a SONY camera in the same room and under constant conditions such as brightness or distance.Regression equation predicting thickness of tongue coating was constructed using binary logistic regression analysis.The optimal cut off of probabilities to diagnosis thick tongue coating was determined by receiver operating curve analysis.χ2 test for paired data and kappa test were used to determine the diagnostic value for recurrence and/or metastasis in colorectal cancer patients.Kolmogorov-Smirnov test was used to determine the distribution of ALT,AST,ALP ALB,TP,GLO,TBIL,DBIL,GGT,LDH,GLU,UA,CA724,CA199,CA242 and CEA.Data with the skewed distribution were presented as median(quartile interval).The association between the thickness of tongue coating and clinical-pathological character was evaluated by chi square test and two-independent-sample test.The two-independent-samplesχ2 was used to determine whether there were significant differences in the thin coating and thick coating between patients with recurrence and/or metastasis and patients without recurrence and/or metastasis.Kaplan-Meier method was used to analyze survival time.Statistical analysis was performed by SPSS(version 16.0).Results:Through retrospective clinical study we found that overall survival of colorectal cancer patients with thick greasy tongue coating is less than the patients with less tongue coating.What’s more,the risk for recurrence or/and metastasis overall survival in thick coating group was higher than thin coating group.In addition,the histological staining of the tongue slices of rats showed that EGFR receptors in the tongue root were the most among whole tongue surface.Tongue thick coating may be due to tumor patients with high levels of serum EGF which results in significantly increasing tongue coating.This finding suggested that the tongue coating of cancer patients may reflect the level of serum EGF levels in patients which may be related to shorter survival time.In addition,another study showed that serum lactic dehydrogenase level in patients with thick tongue coating is higher than patients with thin tongue coating.Conclusions:These studies suggest that tongue coating is likely to reflect some of the growth factor and enzyme levels.By observing the tongue coating we could predict the prognosis of patients and the characteristics of tongue coating may be used as new diagnostic markers to patients with colorectal carcinoma.

Influence of Intravenous Injection of Hydroxyapatite on Mouse Movement Ability

Lactic acid is usually formed during strenuous exercise, and a large amount of lactic acid and slow anaerobic glycolysis in muscle lead to limitations of movement ability. Hydroxyapatite（HAp） is an alkaline inorganic material with a good biocompatibility. It slowly degrades in vivo and releases trace amounts of calcium ions, reducing cell damage by reacting with polylactic acid and neutralizing local acid environment. Whether HAp can reduce the concentration of lactic acid in vivo and improve movement ability is not yet clear. Here, after eight days of training, 40 mice were randomly divided into four groups： control, distilled water, sodium bicarbonate and HAp group. After one day of rest, the mice were intravenously injected via the lateral tail veins with 1 ml distilled water, sodium bicarbonate and HAp suspension（2.2 mg/ml）, respectively, and subjected to a swimming exhaustive experiment（load 10%）. The swimming time of mice in the water, under water and total time were recorded. The exhaustive exercise mice were immediately subject to abdominal cavity anaesthesia. The concentrations of blood, gastrocnemius and myocardial lactic acid as well as serum LDH activity were detected. We demonstrate that HAp can significantly prolong swimming time and improve serum LDH activity, but does not affect lactic acid concentration. In conclusion, intravenous injection of HAp nanoparticles can significantly improve the exhaustive swimming ability of mice mainly because of the elevated blood LDH activity induced by HAp.