OBJECTIVETo study the effects on SOD, MDA, gamma-GT, GSH-Px and inflammatory factor (TNF-alpha, NF-kappaB, ICAM-1) in rats that induced by fructus toosendan, and to search for the hepatotoxicity mechanism of rats that induced by fructus toosendan.

METHODThe SD rats were given fructus toosendan 120 g x kg(-1) by orally for 45 days, then take the liver tissue of control and fructus toosendan group to prepare liver homogenate. The activities of SOD, the content of MDA, the ratio of SOD and MDA, the content of gamma-GT and glutathione peroxidase (GSH-Px) were detected according to the methods of kit. The tumor necosis factor-alpha (TNF-alpha) was detected by ABC-ELISA. The expression of NF-kappaB p65 and ICAM-1 were detected by immunohistochemistry.

RESULTThe rats were given fructus toosendan 120 g x kg(-1) by orally for 45days, the SOD and GSH-Px activities in liver tissue decreased, the content of MDA increased, the ratio of SOD and MDA decreased, the content of gamma-GT and TNF-alpha, the masculine expression of NF-kappaB p65 and ICAM-1 increased.

CONCLUSIONAfter the rats were given fructus toosendan, the liver can be damaged obviously, and the mechanism of hepatotoxicity perhaps related to free radical and inflammatory factor.

Animals ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Female ; Glutathione Peroxidase ; metabolism ; Immunohistochemistry ; Liver ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; NF-kappa B ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; gamma-Glutamyltransferase ; metabolism

AIM To optimize the preparation of two inclusion compounds of erinacine A and to evaluate their stabilities.METHODS With feed ratio,milling time and milling speed as influencing factors,inclusion rate and yield as evaluation indices,the preparation was optimized by orthogonal test.The obtained inclusion compounds were characterized by infrared spectrophotometry and TLC,whose stabilities at high temperature (60 ℃),strong light (3 000 1x) and high humidity [(90 ±5)％] were investigated.RESULTS The optimal β-cyclodextrin inclusion conditions were determined to be 5 ∶ 1 for feed ratio,60 min for milling time,and 300 r/min for milling speed,the inclusion rate and yield were 20.66％ and 88.21％,respectively.The optimal hydroxypropyl-β-cyclodextrin inclusion conditions were determined to be 25 ∶ 1 for feed ratio,90 min for milling time,and 400 r/min for milling speed,the inclusion rate and yield were 69.25％ and 96.31％,respectively.The inclusion of Hericium erinaceus 80％ ethanol extract was physical process without composition change.At high temperature and strong light,two inclusion compounds' appearance showed no obvious change with little loss of erinacine A,which was just the contrary at high humidity.The inclusion effect of hydroxypropyl-β-cyclodextrin inclusion compound was better than that of β-cyclodextrin inclusion compound.CONCLUSION Both β-cyclodextrin and hydroxypropyl-β-cyclodextrin inclusion compounds of erinacine A exhibit good heat stability and light stability,but deliquescence can easily happen to them.

To investigate the effect of treadmill running on the ability of learning in young rats, male Sprague-Dawley rats (5 weeks of age) were used for the experiment. Animals were randomly divided into the control and running groups (n=15 in each group). The rats in running group were made run on a motor-driven treadmill for 1 week at a speed of 2 m/min for the first 5 min, at a speed of 5 m/min for the next 5 min, then at a speed of 8 m/min for the last 20 min. Then the Morris water maze was used to observe learning and memory ability of rats in both groups. The tests consisted of place navigation and spatial probe test. We found that, in place navigation training, the latency of rats in running group was less than that in control group (P<0.05); and from the third training session on, there was significant difference between the rats in control and running groups in swimming velocity (P<0.01); furthermore, it was observed that the rats in running group had stronger motivation and more exact orientation in searching for platform, which could be indicated by the index of turn angle and angular velocity. In spatial probe test, there was no significant difference between the two groups in swimming velocity, percentage of swimming distance and frequency of crossing platform in D quadrant, where the platform situated (P>0.05). These findings suggest that low speed treadmill running can enhance the ability of learning in young rats.
Age Factors ; Animals ; Male ; Maze Learning ; physiology ; Memory ; physiology ; Physical Conditioning, Animal ; physiology ; Rats ; Rats, Sprague-Dawley ; Spatial Behavior ; physiology ; Swimming ; physiology

AIMTo study the expression and regulation of Smad1, Smad2 and Smad4 proteins (intracellular signaling molecules of transforming growth factor-b family) in rat testis during postnatal development.

METHODSThe whole testes were collected from SD rats aged 3, 7, 14, 28 and 90 (adult) days. The cellular localization and developmental changes were examined by immunohistochemistry ABC method with the glucose oxidase-DAB-nickel enhancement technique. Quantitative analysis of the immunostaining was made by the image analysis system. The Smads proteins coexistence in the adult rat testis was tested by the double immune staining for CD14-Smad4 and Smad2-Smad4. The protein expression of Smad during rat testicular development was examined by means of Western blots.

RESULTSSmad1, Smad2 and Smad4 were present throughout testicular development. The immunostaining of Smad1 and Smad2 were present in spermatogenic cells. A positive immunoreactivity was located at the cytoplasm, but the nucleus was negative. Smad1 was immunolocalized at the d14, d28 and adult testes, while Smad2, at the d7, d14, d28 and adult testis. There was positive immunoreaction in the Sertoli cells and Leydig cells as well. The immunolocalization of Smad4 was exclusively at the cytoplasm of Leydig cells and the nuclei were negative throughout the testicular development. No expression was detected in the germ cells. The results of image and statistical analysis showed that generally the expression of Smad1, Smad2 and Smad4 in the testis tended to increase gradually with the growth of the rat.

CONCLUSIONThe present data provide direct evidences for the molecular mechanism of TGF-bgr action in rat testes during postnatal development and spermatogenesis.

Animals ; Blotting, Western ; DNA-Binding Proteins ; analysis ; biosynthesis ; Immunohistochemistry ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology ; Smad Proteins ; Smad1 Protein ; Smad2 Protein ; Smad4 Protein ; Testis ; chemistry ; growth & development ; physiology ; Trans-Activators ; analysis ; biosynthesis

Objective The neuronal ceroid lipofuscinosis (CLN are a group of severe lysosomal storage diseases.The patients present with clinically and genetically heterogeneous neurodegenerative disorders.This study aims to investigate the clinical characteristics and the gene mutations of a rare Chinese family with 3 siblings affected by CLN7.Methods The proband,a 5-year-old girl,visited us because of intermittently seizures and mental retardation for 2 years and a half in December,2015.Clinical investigation,brain magnetic resonance imaging(MRI),biochemical and the gene analysis were performed for the etiological study.Results The proband had seizures at the age of 2 and a half years,with the progressive motor deterioration,speech disturbance,mental regression and vision loss.Her brain MRI showed diffusive cerebral atrophy.The blood aminoacids,acylcarnitine and urine organic acid profiles were normal.Lysosomal palmitoyl protein thioesterase and tripeptidyl peptidase activities of peripheral leukocytes were normal.A compound heterozygous mutation of c.1351-1G ＞ A and c.300T ＞ G was detected on her MFSD8 gene,supporting the diagnosis of CLN7.Both of the 2 mutations were novel.Each of her parents carried one of the mutations.Two brothers of the proband had similar clinical process.Her elder brother died at the age of 7 due to severe encephalopathy of unknown etiology.The younger brother showed dyskinesia from the age of 2 years and seizures from the age of 4 years.A compound heterozygous mutation on MFSD8 gene,c.1351-1G ＞ A and c.300T ＞ G,was found from the younger brother,as same as the proband.Conclusions CLN7 is a rare disorder of CLN.In this study,the diagnosis of the 3 siblings with similar clinical process were much delayed.Gene analysis was key for the diagnosis.Two novel mutations were found on MFSD8 of the family.There is still no effective treatment for neurol ceroid lipofuscinosis.The prognosis is poor.Based on the mutation diagnosis,prenatal diagnosis for the next sibling is possible to the prevention of the disease.

Objective To establish a stable and reliable method for the determination of ethylene oxide residue,and to analyze ethylene oxide residue in multi components made of different materials involved in some medical devices,so as to provide references for sample selection and ethylene oxide residue detection of multi-component medical device kits.Methods A method for the determination of ethylene oxide residue of multi-component medical devices was developed using headspace-gas chromatography and DB-WAX column under the conditions of headspace extraction with equilibration at 80℃ for 20 min,and the weighing mass,linearity,limit of detection,limit of quantification,precision and recovery of the method were determined.Trials of the method were carried out on the items undergoing ethylene oxide sterilization,including disposable perineal care kit,disposable gynecological examination kit,disposable suture dressing kit,disposable debridement kit and the components contacting human body in the disposable dialysis kit,and the abilities of different materials of the components were analyzed in absorbing,retaining and releasing ethylene oxide.Results The method showed high linearity(r=0.999 8)in the range of ethylene oxide mass concentration from 0.4 to 16.0 μg/mL with a weighing mass of 1.00 g,which had the limit of detection being 0.11 μg/mL,the limit of quantification being 0.37 μg/mL and the relative standard deviations(RSDs)for the precision from 0.35％to 1.52％.The average recoveries of different spiked amounts of ethylene oxide in the three blank matrices ranged from 92.68％to 101.42％with the relative standard deviations(RSDs)from 2.46％to 7.59％,which all satisfied the detection requirements.The components made of rubber and acrylonitrile-butadiene-styrene copolymer(ABS)in multi-component medical device kits had the highest ethylene oxide residues,followed by the components made of wood,degreased cotton,polypropylene and polystyrene.Conclusion The method proposed gains advantages in easy operation and high specificity,quantification and reproducibility,which can be used for the determination of ethylene oxide residue in the multi-component medical device kit undergoing ethylene oxide sterilization.References are provided for sample selection of multi-component medical devices.[Chinese Medical Equipment Journal,2024,45(1):56-61]

In this study, an analytical method was developed and used to quantify simultaneously protocatechuic acid, neochlorogenic acid, cryptochlorogenic acid and 1, 3-dicaffeoylquinic acid--four bioactive compounds contained in Fructus Xanthii using UPLC. The contents of four phenolic components of 28 batches of samples collected from different product areas and markets were determined and compared by means of this established method. The mobile phase was composed of methanol and water containing 0.1% phosphoric acid. Chromatography was monitored at dual-wavelengths--220 and 327 nm. Flow rate was 0.4 mL x min(-1) and column temperature was 35 degrees C. The correlation coefficient between concentration and chromatographic peak area of protocatechuic acid, neochlorogenic acid, cryptochlorogenic acid and 1, 3-dicaffeoylquinic acid was over 0.9999 in the range of 0.3570-35.70, 2.500-250.0, 1.060-106.1, 1.010-101.0 microg x mL(-1), respectively. The average recoveries of the four compounds were 97.68%, 99.55%, 97.92% and 100.4%, respectively. In conclusion, the established method can rapidly attain an accurate and reproducible result used to control the quality of Fructus Xanthii.
Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Fruit ; chemistry ; Hydroxybenzoates ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Quinic Acid ; analogs & derivatives ; analysis ; Reproducibility of Results ; Xanthium ; chemistry

OBJECTIVETo evaluate the association between p53 codon 72 polymorphism (R72P) and the risk of colorectal liver metastases.

METHODSThe p53 R72P genotype was identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 78 consecutive colorectal cancer patients with liver metastases and 214 age- and sex-matched cases with nonmetastatic colorectal cancer.

RESULTSThe R allele of the p53 R72P polymorphism was more frequently found in metastatic cases than in nonmetastatic cases (P=0.075). Carriers of the 72R allele had a 2.25-fold (95% CI (confidence interval)=1.05 to approximately 4.83) increased risk of liver metastases. On the stratification analysis, 72R-carrying genotype conferred a 3.46-fold (95% CI=1.02 to approximately 11.72) and a 1.05-fold (95% CI=0.36 to approximately 3.08) increased risk of liver metastases for p53 overexpression-positive and negative colorectal cancers, respectively.

CONCLUSIONThese results demonstrate for the first time that the 72R allele of the p53 polymorphism has an increased risk for liver metastases in colorectal cancers positive for p53 overexpression.

Adenocarcinoma ; genetics ; metabolism ; pathology ; secondary ; Case-Control Studies ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; DNA, Neoplasm ; blood ; genetics ; Female ; Genes, p53 ; Genetic Predisposition to Disease ; Genotype ; Humans ; Liver Neoplasms ; genetics ; metabolism ; secondary ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVETo observe the effect of matrine on human ether à go-go related gene (HERG) potassium channels expressed in Chinese hamster ovary (CHO) cells and investigate whether HERG channel is a new target of the pharmacological effect of matrine on arrhythmia and tumor

METHODSHERG channel potassium current in CHO cell was recorded using whole-cell patch-clamp technique, and the influence of matrine on the current was explored.

RESULTSMatrine inhibited HERG potassium current in a dose-dependent manner, and the 50% inhibitory concentration (IC IC(50)) was 411±23 μmol/L. Matrine had no significant effect on the activation kinetics, and mainly blocked HERG channels in their closed state.

CONCLUSIONSThe blocking effect of matrine on HERG channels might be one of the mechanisms against arrythmias and tumors. Unlike most other blockers exerting blocking effect at the intracellular sites by entering the cell with the opening of HERG channel, matrine blocked HERG channels at the extracellular sites.

Alkaloids ; pharmacology ; Animals ; CHO Cells ; Cricetinae ; Cricetulus ; ERG1 Potassium Channel ; Ether-A-Go-Go Potassium Channels ; genetics ; metabolism ; Humans ; Quinolizines ; pharmacology

OBJECTIVE: To investigate the diagnostic value of diffusion kurtosis imaging (DKI) histogram analysis in hepatic fibrosis staging. MATERIALS AND METHODS: Thirty-six rats were divided into carbon tetrachloride-induced fibrosis groups (6 rats per group for 2, 4, 6, and 8 weeks) and a control group (n = 12). MRI was performed using a 3T scanner. Histograms of DKI were obtained for corrected apparent diffusion (D), kurtosis (K) and apparent diffusion coefficient (ADC). Mean, median, skewness, kurtosis and 25th and 75th percentiles were generated and compared according to the fibrosis stage and inflammatory activity. RESULTS: A total of 35 rats were included, and 12, 5, 5, 6, and 7 rats were diagnosed as F0–F4. The mean, median, 25th and 75th percentiles, kurtosis of D map, median, 25th percentile, skewness of K map, and 75th percentile of ADC map demonstrated significant correlation with fibrosis stage (r = −0.767 to 0.339, p < 0.001 to p = 0.039). The fibrosis score was the independent variable associated with histogram parameters compared with inflammatory activity grade (p < 0.001 to p = 0.041), except the median of K map (p = 0.185). Areas under the receiver operating characteristic curve of D were larger than K and ADC maps in fibrosis staging, although no significant differences existed in pairwise comparisons (p = 0.0512 to p = 0.847). CONCLUSION: Corrected apparent diffusion of DKI histogram analysis provides added value and better diagnostic performance to detect various liver fibrosis stages compared with ADC.
Animals ; Carbon ; Diagnosis* ; Diffusion* ; Fibrosis* ; Liver ; Liver Cirrhosis ; Magnetic Resonance Imaging* ; Rats ; ROC Curve