Aged ; Antibodies, Monoclonal ; metabolism ; Carcinoma, Transitional Cell ; metabolism ; pathology ; surgery ; Cystectomy ; Follow-Up Studies ; Humans ; Keratin-7 ; metabolism ; Keratins ; immunology ; Lung Neoplasms ; secondary ; Male ; Urinary Bladder Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Objective: To optimize the extraction and purification technology of liposoluble constituents from Bufonis Venenum. Methods: The total content of cinobufagin and resibufogenin was taken as the index, and the ethanol potency, solvent multiplication and extraction time were taken as the investigation factors. The optimum extraction process parameters was determined by orthogonal test. Through single factor investigation experiment combined with Box-Behnken response surface method, taking the expansion agent ratio, ratio of diameter to height, adsorbent and sample quantity as the investigation factors, the optimum purification process was selected and determined. Results: It was determined that the optimum extraction process for the addition was 10 times 85% ethanol for 90 minutes’ extraction, the best purification process for the expansion agent cyclohexane-chloroform- acetone was 4:3:3, column height and diameter was 7:1, adsorbent and sample volume was 5.5:1. The content of two kinds of toad poison base purified was up to 66.51%. Through the verification of three batches of amplification process, it was shown that the model fit well, and the separation and purification of toad poison ligand by silicone column chromatography had the advantages of simple operation, fast separation speed, and good separation effect. Conclusion: The selected process is reasonable and feasible, which provides technical reference and basis for the industrialization application of the product in the future.

Objective: The optimum technological conditions of granule dry granulation of Wenshen Zhuanggu Granule (WZG) were optimized to improve production efficiency and ensure product quality and curative effect. Methods: The difficulty degree of granulation was studied by orthogonal test. The molding rate and the dissolved property of the granule determined the type and dosage of excipients for the evaluation index. Through single factor investigation test and combined with Box-behnken response surface method, the feed speed, rolling speed and rolling pressure were the independent variables, and the particle forming rate was dependent variable, in order to optimize process parameters of dry granulation. Results: The optimum proportion of excipients was the dextrin with 1:3 of the dosage of dry paste powder, 1:15 carboxymethy starch sodium, 1:90 of the micro-powder silica gel, the best granulation process for rolling wheel pressure 9.5 MPa, rolling speed 14.0 Hz, and feed speed 13.2 Hz. Through the amplification process validation of three batches, it showed that the WZG prepared by the optimized excipients and process parameters have the advantages of high molding rate, good melting and low moisture absorption rate. Conclusion: The selected process is reasonable and feasible, and it provides technical reference and basis for the industrial application of the product.

Objective To observe the influence of nandrolone phenylpropionate(NP) on the ultrastructure of aorta in rats with or without movement training,and investigate the side effects of NP on the cardiovascular system. Methods Forty male SD rats were randomly divided into sedentary control group,sedentary+medicine group,exercise control group and exercise+medicine group.For the groups with medical treatment,NP of 10 mg/kg one time every three days was injected into the rats via gluteus for eight weeks.For the exercise groups,rats were trained to run on treadmill five days per week for eight weeks.The aortae were sampled and specimens were obtained for transmission electron microscopy. Results The ultrastructure of aorta was normal in sedentary control group.For sedentary+medicine group,mitochondrial swelling,vacuolated cytoplasm and lysis of endothelial cells were observed,disruption of intercellular conjunctions,widening of subendothelial spaces and furcation and breakage of internal elastic lamina were found,and smooth muscle cells changed into synthesis type.For exercise control group,no obvious morphologic change was observed,except that part of the internal elastic lamina disrupted.In exercise+medicine group,breakage and lysis of endothelial cells were observed,widening of subendothelial spaces and lysis of internal elastic lamina were found,and autophagosome and myelinoid body were seen in smooth muscle cells. Conclusion NP may lead to the impairment of endothelial cells and the change of smooth muscle cells into synthesis type.Exercise with NP administration may result in more severe impairment in vessel wall.

Objective To observe the dynamic changes of neuron-specific enolase(NSE)mRNA and protein in offspring rats brain tissue with bilirubin encephalopathy and explore the pathological mechanism and its diagnostic value on bilirubin encephalopathy.Methods Seven-day postnatal Wistar rats were used for study.One hundred and twenty rats were divided into 2 groups randomly(control group and experimental group),which were respectively subdivided into 6 groups(6,12,24,48,72,96 h).The rats in control group were intraperitoneally administered physiological saline 0.5 mL,the rats in experimental groups were intraperitoneally administered bilirubin(200 mg/kg).Reverse transcription-polymerase chain reaction was used to analyze the dynamic changes of NSE mRNA expression at 6,12,24,48,72 and 96 h in brain tissue of rats with bilirubin encephalopathy.Immunohistochemistry was used to evaluate NSE protein expression in hippo-campi,cerebral cortex,thalamic and pallidus at different times.Results The expression of NSE mRNA significantly decreased in brain tissue of rats with bilirubin encephalopathy from 6 h to 96 h compared with the control groups.The expression of NSE protein in hippocampi decreased in offspring rats with bilirubin encephalopathy from 6 h to 96 h,but there were no differences compared with the control groups.The expression of NSE protein in cerebral cortex was significantly decreased in rats with bilirubin encephalopathy from 6 h to 96 h,there were significant differences compared with the control group.The expression of NSE protein in thalamic significantly decreased in rats with bilirubin encephalopathy from 6 h to 96 h,but there were significant differences between experimental groups and the control groups at 24 h and 72 h.The expression of NSE protein in pallidus significantly decreased in offspring rats with bilirubin encephalopathy from 6 h to 96 h,and there were significant differences compared with control groups.Conclusions The changing trends of expression of NSE mRNA were identical to those of NSE protein.NSE may reflect the degree of injury of neurogliocyte.It can serve as reliable index to determine bilirubin encephalopathy.

ObjectiveTo investigate and compare the behavioral changes, neuron loss of hippocampus and mossy fiber sprouting between pilocarpine-induced status epilepticus (SE) model and pentylenetetrazole (PTZ) kindling model in rats.MethodsAfter two different epilepsy models were made, Vedio was adopted to observe the behavioral changes. Nissl staining and Neo-timms' staining were separately used to observe and compare the neuron loss of hippocampus and mossy fiber sprouting in the dentate gyrus (DG) at different time points during epileptogenisis.ResultsNo recurrent spontaneous seizure, no neuron loss and no mossy fiber sprouting were found in PTZ kindling model; whereas obvious neuron loss was found in CA1, CA3 of hippocampus and hilus of DG, and mossy fiber sprouting were found in pilocarpine model in parallel with recurrent spontaneous seizures. ConclusionPTZ kindling model resembles absence epilepsy in human, while pilocarpine-induced status epilepticus model resembles chronic temporal epilepsy in human. Neuron loss and mossy fiber sprouting may play an important role in epileptogenisis. Pilocarpine-induced epilepsy model can be regarded as an ideal chronic temporal epilepsy model.

Syndrome and formulae (or prescription) are two key issues in traditional Chinese medicine (TCM) and the premise research for material basis of TCM. However, vagueness of syndromes and complexity of formulae greatly limited the evaluation to syndromes and effective substance basis of prescription. Therefore, how to solve the evaluation of syndromes, confirming the efficacy material basis in prescription are the current hot issues of international concern. To solve these problems, establishing chinmedomics by integrated serum pharmacochemistry of TCM with metabolomics technology, that is a unique method of TCM research, made outstanding contributions in solving international concerns such as the effectiveness and security aspects of TCM. On the basis of the biological characterization of syndrome, the metabolic profiling of animal models of TCM syndrome, and related metabolic fingerprints as well as metabolic biomarkers were established to evaluate the overall effects of TCM formulae and corresponding relationship of syndrome-formulae. The active constituents were screened using the plotting of correlation between (endogenous) marker metabolites and (exogenous) serum constituents (PCMS), and is ongoing verification by further biological experiments. Correlation analysis between the ingredients in the body after oral formulae and endogenous markers in vivo can be used to clarify the active ingredients and synergistic properties. This method was successfully applied for rapid discovery of potentially bioactive components and metabolites from TCM, and through a series of studies on the chinmedomics, it proved that the established method could help to explore the effective substance for further research of TCM. As a new research approach, Chinmedomics is the best method to fit the holistic concept of TCM, and it can not only interpret the essence of syndrome but also elucidate the scientific connotation of Chinese medical formulae.
Animals ; Diagnosis, Differential ; Drug Prescriptions ; Drug Therapy ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Humans ; Medicine, Chinese Traditional

Objective To analyze the outcome of the epidemiological investigation on an animal plague in Dingbian County of Shaanxi Province.Methods The region of 25 square kiolmeters(km2)in Hongliugou Town was selected as monitoring point of plague to investigate on host animals,etiology and serology.The epidemic area was classified,deratization and depulization were correspondingly adopted.Health education was carried out for prevention knowledge of plague,and questionnaire survey was conducted among residents and medical staff in the epidemic area.Result The average rat density in monitoring site was 8.38 rats per hectare in Hongliugou Town.Average rate infected with flea and flea index were 50.4%(56/111)and 1.81,respectively.The epidemic area was classified 3 types,and came up to the demand after corresponding measures adopted.In the epidemic area,the pass rate of the prevention and control knowledge were 62.00%(31/50)in residents and 92.98%(53/57) in medical staff.Conclusions Plague epizootic can be discovered by exercising regular monitoring and controlledimmed iately and effectively by taking the appropriate control measures.

OBJECTIVETo investigate the pathogenicity of Leptospira interrogans fliR gene to J774A.1 cells.

METHODSfliR gene from L. interrogans serogroup Icterohaemorrhagiae serovar lai strain 56601 and kana gene from plasmid pET42a were amplified by PCR. Suicide plasmid of fliR gene was constructed; and specific siRNA for fliR gene was designed and synthesized. fliR gene mutants were constructed by gene knock-out with suicide plasmid (56601fliR-Kana) and gene silencing with siRNA (56601siRNA-R2). The mutants were identified by PCR, sequencing and semi-quantitative RT-PCR. Adhesion to mouse mononuclear-macrophage J774A.1 and induction of cell necrosis and apoptosis by 56601fliR-Kana and 56601siRNA-R2 were examined by adhesion test and flow cytometry, respectively.

RESULTThe nucleotide and putative amino acid sequences of cloned fliR gene had 99.9% and 100% similarities to those of reported sequences in GenBank. The nucleotide sequence of the cloned kana gene was identical to the corresponding sequence in pET42a map. The results of PCR and sequencing confirmed that kana gene was inserted in the sequence of 56601fliR-Kana fliR gene. The mRNA level of fliR gene in 56601fliR-Kana was remarkably decreased (P<0.01) while the mRNA level of fliR gene in 56601siRNA-R2 was much lower than that in wild strain 56601 (P<0.05). 56601fliR-Kana and 56601siRNA-R2 lost the ability to adhere J774A.1 cells; and their ability to induce cell necrosis and apoptosis was markedly weakened (P<0.01).

CONCLUSIONfliR is a virulence-associated gene of L. interrogans and the function of the gene is closely related to adhesion, induction of cell necrosis and apoptosis of the microbe.

Animals ; Apoptosis ; Bacterial Adhesion ; Bacterial Proteins ; genetics ; metabolism ; Cell Line ; Leptospira interrogans ; genetics ; pathogenicity ; Macrophages ; microbiology ; pathology ; Membrane Proteins ; genetics ; metabolism ; Mice ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics

0.05),the mean FA on the left was higher than the right(t=1.912,P