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Objective:To investigate the expression of circ_0063865 in esophageal squamous cell carcinoma(ESCC)tissues and cells and its effect on the biological properties of the cells.Methods:The loop structure and stability of circ_0063865 were identified by Sanger sequencing, back-to-back primer validation and the ribonuclease R(Rnase R)tolerance assay.The expression of circ_0063865 was detected by RNA fluorescence in situ hybridization in an ESCC tissue microarray and its clinical relevance was analyzed.The expression levels of circ_0063865 in a normal esophageal epithelial cell line and ESCC cell lines were measured by real-time quantitative polymerase chain reaction(RT-qPCR). Cell counting Kit-8, the colony formation assay, the scratch assay, the transwell invasion assay and flow cytometry were used to detect the effects of circ_0063865 on cell proliferation, migration, invasion abilities and apoptosis, respectively.Results:The loop formation of circ_0063865 was verified by Sanger sequencing, back-to-back primer and Rnase R tolerance assays.The results of RNA fluorescence in situ hybridization showed that the mean fluorescence intensity of circ_0063865 expressed in ESCC tissues was significantly higher than in its paired paracancerous normal tissues( t=2.267, P<0.05). The expression of circ_0063865 was significantly associated with lymph node metastasis( χ2=4.356, P<0.05). The average overall survival time of patients with high circ_0063865 expression ESCC was lower than that of patients with low circ_0063865 expression ESCC.RT-qPCR results demonstrated that, compared with HEEC, circ_0063865 expression was elevated in ESCC cell lines( F=18.413, P<0.05). In addition, after circ_0063865 knockdown, the proliferation, migration and invasion abilities of KYSE-30 and KYSE-150 cells were significantly decreased, and the level of apoptosis was significantly increased(both P<0.05). Conclusions:The expression of circ_0063865 in ESCC is high, and changes in its expression are significantly correlated with lymph node metastasis.Additionally, circ_0063865 can promote the proliferation, migration and invasion of ESCC cells.
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With the development of global economy, the dramatically increased production of polyethylene terephthalate (PET) plastics has led to a remarkably increased amount of plastic waste. PET waste can be treated by landfill, incineration, or biodegradation. While landfilling and incineration may cause secondary pollution, biodegradation has since received increased attentions due to its environmental friendliness. Recent studies have indicated that the carbohydrate binding module (CBM) can effectively enhance the binding of PET degrading enzymes to PET, and consequently increasing PET degradation rate. Here we constructed a fusion protein BaCBM2-Tfuc containing the BaCBM2 from Bacillus anthraci and the cutinase Tfuc from Thermobifida fusca, by megaprimer PCR of whole plasmids (MEGAWHOP). Notabaly, the PET film degradation efficiency (at 60 ℃) of BaCBM2-Tfuc was 2.8 times that of Tfuc. This study may provide technical support for constructing fusion proteins capable of efficiently degrading PET.
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Carbohydrates , Carboxylic Ester Hydrolases , Polyethylene Terephthalates , ThermobifidaABSTRACT
Objective To explore the clinical features and gene mutations of blepharophimosis-ptosis-intellectual-disability syndrome (BPID). Methods The clinical data, diagnosis and treatment of a child with BPID in neonatal intensive care unit (NICU) were reviewed. Based on the literature retrieved from PubMed database, the common classification, clinical features, diagnosis and genetic counseling of BPID and its affiliated blepharophimosis-mental retardation syndromes (BMR) were reviewed. Results This male infant was 39 weeks of gestational age with birth weight of 1920 g, and was admitted to NICU 15 min after birth due to dyspnea. The main clinical manifestations were facial deformity such as biepharophimosis, ptosis and micromandible, inspiratory dyspnea with laryngeal cartilage softening, malformations of the thorax and feeding difficulties. A heterozygous mutation in UBE3B gene was identified by complete exon sequencing and he was diagnosed of BPID, a rare genetic disorder. Reviewing the literature, there was no relevant report in domestic. While one foreign literature was found to report 5 patients from 4 families having a subtype of BMR, a kind of autosomal recessive diseases caused by mutations in the UBE3B gene. Conclusion BPID is a rare clinical entity of BMR. Complete exon sequencing can be used to diagnose the disease.
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Objective To develop a new risk model for predicting type 2 diabetes (T2D) in a rural Chinese population in north China.Methods A village-based cohort study was performed.Data from Handan Eye Study conducted from 2006-2013 comprising 4 132 participants aged 30 years old (1 793 male and 2 339 female) with complete diabetes data at baseline and follow-up were analyzed.The blood biomarkers of T2D incident risk were screened and a new risk model was derived by using unconditional stepwise logistic regression after adjustment of age,body mass index (BMI),waist circumference,and family history of diabetes in random two-thirds of the sample cohort (selected randomly).In addition,a simple point system for T2D risk was built according to the procedures as described in Framingham Study,and the new risk score was subsequently validated in the final one-third of the sample cohort.Results The new risk score included age (8 points),BMI (6 points),waist circumference (8 points),family history of diabetes (9 points),fasting plasma glucose (23 points),and triglycerides (4 points).The score ranged from 0 to 58.The AUC was 0.802 (0.780-0.822) in the validation sample.At the optimal cutoff value of 27,the sensitivity and specificity were 70.27％ (58.50％-80.30％) and 80.83％ (78.60％-82.90％) respectively.Conclusions A new risk model for predicting T2D have been developed in a rural Chinese population in north China,and the risk score can be used in rural basic health care settings after validation.
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Gastrointestinal symptoms such as regurgitation,infantile colic,constipation,dyschezia,and diarrhoea are frequent in infants.These symptoms always were functional gastrointestinal symptoms.It is easily confused with other diseases.The knowledge and management of these symptoms are no uniform or clear.Since double-blind placebo controlled prospective intervention trials are very limited,the algorithms are still based on consensus using the evidence.This article respectively introduces each gastrointestinal symptoms,include clinical manifestations,diagnosis and management referring to Rome Ⅳ and domestic and overseas paediatricians consensus,to improve clinical pediatricians the knowledge of gastrointestinal symptoms and specify the management.
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Objective To analyze the differences of biochemical and immunological indicators in gender or Lee′s classification of IgA nephropathy ( IgAN) to provide laboratory evidence for clinical diagnosis and treatment of IgAN.Methods Retrospective cohort study.The information of biochemical and immunological indicators of 213 in-hospital patients which were admitted in Chinese PLA General Hospital in June to December, 2012.The data were collected and analyzed with t-test, non-parameter analysis, Pearson correlation analysis, ROC curves analysis and Logistic regression analysis according to gender or Lee′s classification ( Lee′s≤3 group and Lee′s>3 group).Results In this study, the average age and sex ratio of patients with IgAN was ( 35.0 ±10.6 ) years old and 2.04∶1.00.T-test and non-parameter analysis indicated homocysteine(HCY), immunoglobulin E (IgE), immunoglobulin M (IgM), prealbumin (PA), ceruloplasmin ( CP) ,α1-acidoglycoprotein(α1-AGP) , albumin ( Alb) ,α2-globulin(α2-G) andγ-globulin (γ-G) had significant difference in gender (P<0.05).And cystantin-C (CYS-C), HCY, complement 4 (C4),β2-microglobulin(β2-MG), PA, α1-AGP, α1-globulin(α1-G), immunoglobulin G (IgG), IgM and transferrin ( TF ) had significantly different in Lee′s classification ( P<0.05 ).ROC curves analysis showed that the areas under curves of CYS-C andβ2-MG were 0.891 and 0.839 respectively;the sensitivity was 90.2%and specificity was 75.3% when cutoff value of CYS-C was 1.56 mg/L; the sensitivity was 80.3%and specificity was 78.8% when cutoff value of β2-MG was 0.275 mg/dl.Logistic regression analysis showed that CYS-C ( OR: 31.380, 95% CI: 10.808 -91.113, P=0.000 ) and HCY ( OR:1.035, 95%CI:1.002-1.069, P=0.040) were introduced into the model after twice variable selections.ROC curve of CYS-C combined with HCY for classifying Lee′s classification showed that the sensitivity, specificity, Jorden index and AUC were 0.843, 0.862, 0.705 and 0.894 ( P=0.000 ) , respectively.Conclusions CYS-C,β2-MG, PA and HCY were valuable indicators for Lee′s classification.CYS-C andβ2-MG had high correlation with Lee′s classification, which indicated that these two tests were related to the severity of IgAN.CYS-C combined with HCY could improve the diagnostic efficiency of IgAN Lee′s classification.Combination of biochemical and immunological indicators could improve the accuracy of Lee′s classification and provide effective laboratory evidence for clinical treatment of IgAN.
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Objective To investigate the harmless biochemical indicators for early screening of populations at high risk of suicide .Methods Seven hundred and twenty-seven new recruits were investigated according to self-rating idea of suicide scale (SIOSS) in the first month of recruitment in 2014.Twenty-five recruits (originally 27 recruits, but 2 recruits were absent from biochemical tests ) whose SIOSS scores were less than 12 were recognized as the suicidal ideation group. Another 25 recruits whose SIOSS scores were more than 12 were randomly selected as the control group .Saliva samples of the 50 recruits were collected for biochemical analysis after one month and three months of training , respectively .After three months of training , the SIOSS was also used for evaluation of suicidal ideation .Results After one-month training , the concentrations of Ca , Mg, amylase ( Amy ) and salivary acid ( SA ) in saliva were demonstrated to be statistically different between the suicidal ideation group and the control group ( P 0.05).After three-month training, the concentrations of Ca, Mg, Amy and SA in saliva between the two groups showed no significant difference (P>0.05).Conclusion After the first month training , changes in the biochemical compositions were found in the saliva of the screened recruits with suicidal ideation, which may be associated with the stress response of the body .The biochemical indicators in saliva lack specificity for suicidal ideation .
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Objective:To study the clinical and pathological features of Langerhans cell histiocytosis (LCH), and to provide the reference for its diagnosis and treatment.Methods:The manifestation of one LCH patient was retrospectively analyzed.The features of the LCH patients were explored by analyzing the results of skin biopsy, radiological test and follow-up.The associated literatures were reviewed.Results:The patient presented the typical symptoms gradually,including polycystic lung,skin ulcer,diabetes insipidus,and lactation.The skin pathological findings showed the densely distributed mononuclear cell infiltration in dermis and the immunohistological staining result showed positive CD1a. The patient was follwed up for 7 years and died of heart and lung failure. Conclusion:LHC has various manifestations and should be confirmed by clinical features,pathological features and imaging examination.The adult patients with multisystem and vital organ involvement suggest the poor prognosis.
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Objective:To establish a determination method for sulfadoxine, sulfathiazole and sulfadiazine in animal derived prod-ucts by ultra-performance liquid chromatography-tandem mass spectrometry ( UPLC-MS/MS) . Methods:Trichloroacetic acid and ace-tonitrile were used in the extraction and removal of proteins. A Waters Acquity UPLC BEH C18 (100 mm × 2. 1 mm,1. 7 μm) column was used as the separation column. The mobile phase was 0. 1% formic acid in water-methanol with gradient elution. The three sulfona-mides residues were detected with multiple reaction monitoring( MRM) mode in multi-period. Results:Under the conditions,the linear range of sulfadoxine, sulfathiazole and sulfadiazine was 1. 0-200. 0 ng·ml-1 ,the linear correlation coefficient was above 0. 99,the re-covery was more than 70%,the detection limit was 0. 5 ng·ml-1 , and the quantification limit was 1. 0 ng·ml-1 . Conclusion: The sample preparation method is simple and fast, and the method can be used to analyze sulfadoxine, sulfathiazole and sulfadiazine in ani-mal derived food efficiently and sensitively.
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Objective To compare the two kinds of purification method for purifying recombinant human cardiac troponin I(cT-nI)to obtain the stable cTnI and promote the study of cTnI diagnosis standardization.Methods The cTnI inclusion body was ob-tained by the ultrasonic broken engineering,after washing by 2% Tritonx-100,2M urea,dissolved in 8M urea,then purified by the column refolding on CM-FF and the dilution refolding respectively.The cTnI yields were compared between the two kinds of meth-od and the stability at 4 ℃,20 ℃,-80 ℃ and on the freeze-dried condition was compared.Then the purification method to effi-ciently obtain the stable cTnI was established.Results The protein about 2 mg and 1.4 mg could be obtained by CM-FF on the col-umn refolding and the dilution refolding from 0.1 g of wet inclusion body,respectively.The former method had the short cycle and high efficiency.The cTnI purified by the column refolding on CM-FF was more stable at 4 ℃,20 ℃,-80 ℃ and on the freeze-dried condition.Conclusion The column refolding on CM-FF is more stable and highly efficient in purification of cTnI than the dilution refolding.
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Objective To prepare and identify the monoclonal antibody against asymmetric dimethylarginine (ADMA) and to do further research on clinical application .Methods The short immune peptide antigen was synthetized by solid phase technology . Splenocytes from the Balb /c mice immunized by synthetized antigen were fused with myeloma cells SP 2/0 for producing hybrido-ma .Hybridoma cell line secreting antibodies against ADMA was sifted by indirect ELISA and limiting dilution assay .The specificity of monoclonal antibodies against human ADMA were evaluated with Western blot and ELISA .Results Two hybridoma cell lines stably secreting monoclonal antibodies against ADMA were developed and named 22-1-1and 38-1-6 respectively .By applying West-ern blot and ELISA ,the results indicated that all monoclonal antibodies raised could specifically react with ADMA .Conclusion The success in the production of ADMA monoclonal antibody establishes foundation for application in the practice of clinical labora-to ry .
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Glyeated hemoglobin is a widely used important test in the diagnosis and management of Diabetes Mellitus.Although the Reference Measurement System,Traceability and Clinical for HbA1c have reached international consensus,there are still quite a few aspects related with its clinical and laboratory measurement needed further research.
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Objective To develop a sensitive, simple, and accurate method for the determination of shionone in rat plasma after ig administration of Asteris Radix petroleum ether extract (RAPE). Methods The separation was achieved by HPLC on a RP18 column (150 mm × 3.9 mm, 5μm) with a mobile phase composed of acetotitrile-0.05% phosphoric acid water (98: 2) at a flow rate of 1.0 mL/min. UV Detector was set at 200 nm and friedelin was chosen as an internal standard. Results The linear range of the standard curves was (0.3443-22.0) μg/mL with the correlation coefficient of 0.9968. The intra- and inter-day precisions were all below 10% and the relative error was -3.5%-1.1%.Conclusion The developed method can be successfully applied to the pharmacokinetic study. After ig administration of RAPE, T1/2(ka) is (33.09 ± 7.32) min and T1/2(ke) is (84.95 ± 22.34) min.
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Objective To evaluate the efficacy of long-pulsed tunable Nd: YAG laser on capillary hemangioma and to analyse its influencial factors. Methods A retrospective review was conducted on 124 patients with 135 vascular lesions treated with long-pulsed tunable Nd:YAG laser from February, 2007 to January, 2008. Working parameters, including spot size (3-5 mm), pulse duration (10-30 ms) and fluence (120-200 J/cm2) were determined based on the size, color and depth of lesions and adjusted according to the immediate reaction of lesions. The interval of two treatments was 1 month. Results Of the 135 lesions, 100 experienced a more than 95% subsidence, 22 achieved 60%-94% subsidence, 11 got 30% -59% subsidence, and 2 reached less than 30% subsidence. The cure rate and effective rate were 74.07% and 90.37% respectively. Smaller lesions showed a better response to the treatment than larger ones (P<0.0001), whereas the sites and depth of lesions had no effect on the efficacy (both P>0.05). Atrophy scar occurred in 4 (2.96%) lesions, and blisters in 12 (8.89%) lesions after the treatment. No purpura or pigmentary changes occurred. Conclusion The long-pulsed tunable Nd:YAG laser is an effective treatmnt for capillary hemangioma with fast action and few side reactions.
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Objective To study the stability and reproducibility in serum proteome by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS).Methods Bruker Daltonics MALDI-TOF-MS was adopted under the linear mode, with mass ranging from 800 to 10 000 m/z.MB-WCX pre-extraction kit and Clinprotools software was used to analyze data.Results The reproducibility study was carried out under the standardized conditions.Data analysis showed that the m/z ratio was in the acceptable range, and the coefficients of variation(CV) was 12.7% ,13.1% and 18.8% ,respectively in the parallel experiment by taking 2 cancer and 1 healthy control serum samples and could identify the diseased samples.Then 30 healthy control samples were used to validate this result, all of which showed very good mass profile. The average CV was 25.5%, which could meet the requirements of clinical laboratory.Conclusion With the help of optimized experimental conditions and MALDI-TOF-MS, stable and reproducible mass spectrum can be required which is helpful for the early diagnosis of diseases.
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Objective To develop a candidate reference method for the determination of sodium in serum by inductively coupled plasma mass spectrometry method (ICP-MS). Methods Aluminum, as internal standard of sodium, was added into serum samples and sodium standard solutions by gravimetric analysis. The samples were digested by HNO3 and diluted, and its 23Na/27Al isotope ratios were obtained by ICP-MS. The sodium concentrations were calculated with the standard curve method in serum. Results The analytical recoveries of sodium were 100.67% and 100.15% respectively, and the precisions were 0.08% and 0.04% respectively for 2 different serum samples. The results of measuring sodium in serum of Standard Reference Material (SRM) gave the coefficients of variation (CVs) of 0.18% and 0.22% for 2 levels of standard reference material(SRM) 909b and 0.41%, 0.41% and 0.66% for 3 levels of SRM 956b. The relative deviations between the results and median of the certified value were 0.17% and 0.14% for 909b and -0.09%, -1.05%, -0.48% for 956b respectively. Conclusions The ICP-MS and aluminum internal standard method is proved to be not only precise and accurate, but also quick and convenient for measuring sodium in serum. It is promising to be a candidate reference method for determination of sodium in serum.
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Objective The present study was designed to investigate changes in serum or plasma concentrations of nitric oxide and its derivatives in diabetic patients.Methods Serum nitrate concentration of 84 diabetic patients was measured by using an enzyme kinetic method,and the plasma S-nitrosothiols concentration of 10 cases was measured by using HPLC technique.Results Serum nitrate concentration and plasma S-nitrosothiols concentration in the diabetics were significantly higher than in control group (P＜0.01 andP＜0.05,respectively).The serum nitrate concentration in diabetics also had a significant positive correlation with the serum glucose concentration (R=0.7256,P＜0.05),but this correlation was not found in control group.Conclusion These data showed that NO and its derivatives are overproduced in the diabetic patients.(J Geriatr Cardiol 2008;5:25-27)
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BACKGROUND: Puerarin possesses various biological efficacies, such as the protective efficacy on hypertension, cardiac disease, diabetes mellitus and blood disease, and the extracts of puerarin can inhibit the proliferation of S180 sarcoma and Lewis lung cancer to some extent.OBJECTIVE: To investigate the modulation effects of total flavone of puerarin (TFP) on the growth of pheochromocytoma PC12 cells and the protective efficacy on the H2O2-induced cellular oxidative damage.DESIGN: Complete randomization design and control experiment.SETTING: Department of Biochemistry and Institute of Geriatrics, Chinese PLA General Hospital.MATERIALS: Puerarin was bought from Tongrentang drug store, and TFP was extracted and purified routinely by ethanol and ether acetate, then was evaluated with thin-layer chromatography. The content of puerarin in extracts was 31.79% in quantitative assay. Methyl thiazolyl tetrazolium (MTT), luminal and anti-oxidative activity reagent xanthine oxidase were all from Sigma Company. PC12 cells were given by Institute of Geriatrics,Chinese PLA General Hospital as a present.METHODS: The experiment was conducted at the Institute of Geriatrics, Chinese PLA General Hospital from January to July in·2001.① The cells were cultured in 20 g/L DMEM (pH 7.1-7.2), and randomized into two groups: TFP group and H2O2-injury+TFP group, and each group was divided into 5 mass concentrations (0, 1.0, 10, 100 mg/L and 1.0 g/L). There were 8 holes for parallel culture with 100 mL culture medium in each hole (containing 1 ×l09 L-1 cells). TFP group:TFP was added for 72-hour culture at 37 ℃; H2O2 injury+TFP group:TFP was firstly cultured for 48 hours at 37 ℃, then 500 mmol/L H2O2 was added and co-cultured for other 24 hours.②The activity of cultured PC12 cells was monitored by MTF assays, the content of nitrite was measured by Griess reagents, and the antioxidant activity of superoxidedismutase (SOD) was monitored by hypoxanthine/xanthine oxidase.H2O2-initiated PC12 cellular oxidative damage had been used as experimental model to study the protective efficacy of TFP, and expressed as inhibition ratio [(blank A value-detection A value)/blank A value × 100%]. The higher inhibition ratio indicated the strong ability of clearing O2-. ③ One-factor analysis of variance was used to compare the difference between data. MAIN OUTCOME MEASURES: The influence of TFP on the nitrite content, SOD activity and cell activity in PC12 cells.RESULTS: ①Effect of TFP on PC12 cell activity: 1-10 mg/L TFP hadno obvious effects on the growth of PC12 cells, and 100 mg/L TFP in creased the cell growth (P ＜ 0.05), whereas the TFP concentration was increased to 1.0 g/L, the activity of PC12 cells was inhibited obviously (P ＜ 0.01). TFP of 1-100 mg/L could protect the cultured cells from the oxidative damages of H2O2 concentration dependently (P ＜ 0.05).② Effect of TFP on clearing O2-: The ability of clearing O2 increased withthe mass concentrations of TFP in both groups with obvious dose-effect relation, except when 1 mg/L TFP was added in the H2O2 injury+TFP group. The SOD activity in PC12 cell culture liquid was obviously en hanced after adding 100 mg/L and 1 g/L TFP, compared with that without TFP addition (P ＜ 0.05-0.01). ③Modulation of TFP on nitrite: TFP of low concentration (1-100 mg/L) reduced the production of cellnitrite, whereas increased the nitrite production at the concentration of1.0 mg/mL.CONCLUSION: ①TFP can regulate the growth of PC12 cells, which canbe enhanced by low-concentration (1-100 mg/L) TFP whereas inhibited byhigh-concentration (1 g/L) TFP. However, the anti-oxidation of TFP is themost powerful. ②TFP can protect the PC12 cells obviously from the oxidative damages induced by H2O2 at low concentration.
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BACKGROUND: Disturbance of lipid metabolism is responsible for many kinds of diseases, and to screen safe and effective prescription for lipid reduction has clinical significance.OBJECTIVE: To study the influence of puerarin-arginine composition on lipid metabolism in rats.DESIGN: Comparative observing study based on animal model.SETTING: Experimental animal center and department of biochemistry in a military hospital of Chinese PLA.MATERIALS: This study was carried out at the Experimental Animal Center and Department of Biochemistry of the General Military Hospital of Chinese PLA between May and August 2004. Eighty Wistar rats were randomly divided into high dosage, middle dosage, low dosage group and control group with 10 male and 10 female rats each.METHODS: Rats took food freely, and experimental groups were supplemented with puerarine-arginine composition of different dosage (0, 0.93,1.39 and 1.85 g/kg) for observing the influence on rats.MAIN OUTCOME MEASURES:① blood parameters; ② change of body mass and utilization rate of food; ③ parameters of vital organs;④histopathological changes.RESULTS: From onset of the second week, body mass of both male and female rats in experimental groups were found obviously lower than those in control group. The levels of blood cholesterol in female rats were(1.63 ±0.20) and (1.29 ±0.23) mmol/L in 0.93 g/kg dosage group and control group respectively, and were(1.75 ±0. 13) and(1.31 ±0. 19) mmol/L in male rats,and both have statistical significance( F = 6.43, 6. 58, P ＜ 0. 05).Fatty parameters of the three experimental groups were found obviously lower as comparing with those of control group.CONCLUSION: Such composition has been proved to be able to exert function in lipid reduction by decreasing body mass, and improving utilization rate of food and proportion of cholesterol and organic fat.
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The latest advancement on tumor biology and tumorigenesis has been discussed in this paper. The potential clinical significance on tumor tendency and early diagnosis of the genomic and proteomic analysis technology has been expounded too. It will be benefit for the new tumor biomarker studies.