The present study aimed to investigate the effect and the mechanisms of Bu-zhong-yi-qi-tang (BZYQ) on Spleen-Qi deficiency rat's model using nuclear magnetic resonance (NMR) metabolomics and multivariate statistical analysis methods. The rat Spleen-Qi deficiency model was established as follows: oral administration of Radix Rhei extract, loaded swimming and starvation for 24 h. The body weight and motor behavior of the rats were measured and recorded once a week. BZYQ could significantly improve body weight and behavioral of Spleen-Qi deficiency model rats compared with the model group (P < 0.05, 0.01). After drug administration, the changes in the levels of endogenous metabolites in the spleen including decreasing lactate, taurine and hypoxanthine, increasing glutamate and scyllo-inositol compared with the model group. The metabolomics approach is an effective tool for the investigation of the pharmacologic mechanism of BZYQ and it is helpful to further research.
Administration, Oral ; Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Medicine, Chinese Traditional ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Qi ; Rats ; Spleen

OBJECTIVETo investigate if the interaction between TGF-beta1/Smad pathway and ERK pathway in vascular smooth muscle cells exists.

METHODSThe rat arota was removed. The primary VSMC were isolated and cultured in vitro, then the VSMC were divided into four groups: (1) control group, (2) (TGF-beta1 group, (3) ERK blocking agent group, (4) TGF-beta1 + ERK blocking agent group. The expression of Smad2/3, ERK1/2 proteins, the content of phosphorylated ERK1/2 and Smad2/3 proteins were detected by Western blot, and the expression of Smad2/3 mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR) .

RESULTS(1) In contrast to control group, the content of phosphorylated Smad2/3 and phosphorylated ERK1/2 proteins in TGF-beta1 group was increased (P < 0.05), that in ERK blocking agent group was decreased (P < 0.05). There was no difference between control group and TGF-beta1 + ERK blocking agent group. Compared with TGF-beta1 group, the contents of phosphorylated Smad2/3 and phosphorylated ERK1/2 proteins in TGF-beta1 + ERK blocking agent group was decreased (P < 0.05). There was no difference in the expression of Smad2/3 and ERK1/2 proteins among different groups. (2) There were no differences in expression of Smad2 and Smad3 mRNA among different groups.

CONCLUSION(1) TGF-beta1 can induce Smad2/3 proteins to be phosphorylated dependent on the activated ERK pathway. (2) ERK pathway does not effect the expression of Smad2/3 at the level of protein and mRNA.

Animals ; Aorta ; cytology ; Cells, Cultured ; Female ; MAP Kinase Signaling System ; physiology ; Male ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; physiology ; Myocytes, Smooth Muscle ; physiology ; Phosphorylation ; Rats ; Rats, Wistar ; Signal Transduction ; Smad Proteins ; metabolism ; physiology ; Transforming Growth Factor beta1 ; physiology

OBJECTIVETo establish a high-sensitivity, high-specificity and low-cost hydrogel chip platform for the clinical screening of Y chromosome microdeletions.

METHODSSite-specific extended primers with a common sequence at the 5' end were used for hybridizing with the target. The Cy5-dUTP was incorporated into the products by primer extension, and the products were labeled with fluorescence. Then the extended products were added to the chip for hybridizing with acrylamide-modified common probes immobilized on the chip. After removal of the free Cy5-dUTP by electrophoresis, the signals were obtained by fluorescence scanning. And the detecting conditions of this method were optimized.

RESULTSSY254 of 9 samples was successfully detected with the hydrogel chip. The results showed that 3 were normal and the other 6 with microdeletions (1 female sample as a negative control), which coincided with the results of conventional multiplex PCR-electrophoresis.

CONCLUSIONThe hydrogel chip platform we established has provided a new technique for the detection of Y chromosome microdeletions, and is beneficial to the diagnosis and treatment of male infertility.

Carbocyanines ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Deoxyuracil Nucleotides ; Humans ; Hydrogels ; Infertility, Male ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; diagnosis ; genetics

Objective To investigate the expressions of human telomerase reverse transcriptase(hTERT) mRNA and protein in pheochromocytoma and paraganglioma and their significance as diagnostic markers in predicting the biological behaviour of these tumours.Methods Expression of hTERT mRNA was determined by in situ hybridization in 45 pheochromocytomas/paragangliomas(31 benign,7 suspected malignant and 7 malignant) and 9 normal adrenal medulla samples,hTERT protein was determined by immunohistoebemistry.Results hTERT mRNA was expressed in 5/7 malignant turnouts and 5/7 suspected malignant tumours as compared with 3/31 benign tumours(P

OBJECTIVETo establish a model of acoustically evoked short latency negative response (ASNR) in guinea pigs, a model of profound hearing loss with normal saccular functions, and verify the correlation between ASNR and vestibular evoked myogenic potential (VEMP).

METHODSThirty-two healthy guinea pigs were employed in the experiment, which were randomly divided into control group (16 subjects) and deafened group (16 subjects). Each animal experienced auditory and vestibular tests including auditory brainstem response (ABR), VEMP and caloric test. A quick treatment was employed for deafened group consisting of a subcutaneous injection of kanamycin at a dose of 400 mg/kg followed by a jugular vein injection of ethacrynic acid at a dose of 40 mg/kg one hour later. The animals were received ABR, VEMP and caloric test 7 - 10 days following the drug administration. The deafened group was further divided into ASNR group and non-ASNR group, based on the presence of ASNR.

RESULTSIn deafened group, five subjects died postoperatively, 11 subjects (22 ears) provided full data, ASNR was elicited in eight ears (36.4%), the threshold was 120 - 130 dB SPL with mean of (124.4 ± 4.96) dB SPL. Its latency range was 1.75 - 2.60 ms with mean of (2.15 ± 0.27) ms. The mean latency of threshold was (2.34 ± 0.18) ms. All eight ASNR ears presented with VEMP. The VEMP threshold, positive and negative potential latencies proved no statistical difference (P > 0.05) between ASNR group and control group. Significant difference was detected between the VEMP presence of ASNR group and non-ASNR group (P = 0.002). There was no statistically significant correlation between VEMP and caloric test neither between ASNR and caloric test in deafened group.

CONCLUSIONSThis study evoked ASNR in an ototoxicity guinea pig model which has profound hearing loss with normal saccular functions. The presence of ASNR correlated with VEMP, however, not correlated with caloric test, suggesting that ASNR and VEMP are both originated from the saccule.

Animals ; Deafness ; physiopathology ; Disease Models, Animal ; Evoked Potentials, Auditory ; physiology ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Saccule and Utricle ; physiology ; Vestibular Evoked Myogenic Potentials ; Vestibular Function Tests

OBJECTIVETo establish a model of ototoxicity in guinea pigs with acoustically evoked short latency negative response (ASNR) and verify the responsible organ of ASNR based on microscopic characteristics of basal membranes, saccules, utricles and ampulla canalis semicircularis of the inner ear.

METHODSTotal of 45 guinea pigs were employed in the experiment, which were randomly divided into the control group (15 subjects, 30 ears) and the deafened group (30 subjects, 60 ears). Each animal experienced auditory brainstem response (ABR). A quick treatment was employed for deafened group consisting of a subcutaneous injection of kanamycin at a dose of 400 mg/kg followed by jugular vein injection of ethacrynic acid at a dose of 40 mg/kg one hour later. The animals were performed ABR test from 7 to 10 days after the drug administration. The deafened group was further divided into ASNR group and non-ASNR group based on the presence of ASNR. All the guinea pigs were sacrificed after ABR tests. The Corti organ, macula sacculi, macula utriculi and crista ampullaris were observed by light microscope.

RESULTSIn the deafened group (60 ears), 3 subjects died postoperatively, 27 subjects (54 ears) provided full data. ASNR was elicited in 19 ears (35.2%, 19/54), the thresholds of ASNR were from 110 to 125 dBSPL with average of (121.7 ± 4.5) dBSPL. ASNR latency ranges were 1.80 - 2.08 ms, the average latency of thresholds were (1.93 ± 0.07) ms. The stretched preparation results: overall hair-cell density of macula saccule, macula utriculi and crista ampullaris decreased in order of normal control group, ASNR group and non-ASNR group. There was no difference between the normal group and ASNR group for cell density of macula saccule. Apart from this, statistical differences were found among other groups.

CONCLUSIONSThe present study evoked ASNR in an ototoxicity guinea pig model which was profound hearing loss with normal saccular function and normal saccular hair cell density. It suggested that ASNR originates from the saccule and have no relation with cochlear, utricle and semicircular canal according to morphological study.

Acoustic Stimulation ; Animals ; Deafness ; physiopathology ; Ear, Inner ; physiopathology ; Evoked Potentials, Auditory ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Reaction Time ; Saccule and Utricle ; physiopathology

Gene Expression ; Hep G2 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Liver Neoplasms ; genetics ; metabolism

OBJECTIVETo study the correlation between high-resolution HLA-A * 1101, HLA-A * 0201, HLA-A * 2402, HLA-B * 4001, HLA-DRB1 * 0901 with HCMV pp65 antigenemia after bone marrow transplantation (BMT) in China.

METHODS48 recipients doing BMT during 2009. 2-2010. 10 were selected in my hospital; HCMV pp65 was detected by ELISA or immunohistochemical methods. The frequency of HLA-A * 1101, HLA-A * 0201, HLA-A * 2402, HLA-B * 4001, HLA-DRB1 * 0901 alleles were determined by Polymerase chain reaction-sequence based typing (PCR - SBT).

RESULTS(1) The BMT recipients were HCMV pp65 antigenic positive(100%); (2) The positive rate of HLA-A * 1101, HLA-A * 0201, HLA-A * 2402, HLA-B * 4001 showed no obvious difference between 12 lower antigenemia group and 36 higher antigenemia group, the positive rate: HLA-A * 1101 were 33.3% (8/24) and 20.8% (15/72), HLA-A * 0201 were 4.2% (1/24) and 13.9% (10/72), HLA-A * 2402 were 12.5% (3/24) and 19.4% (14/72), HLA-B* 4001 were 16.7% (4/24) and 12.5% (9/72); (3) HLA-DRB1 * 0901 positive rate in higher antigenemia group was higher than the lower (P = 0.048), the positive rate were 4.2% (1/24) and 19.4% (14/72); (4) HLA-DRB1 * 0901 recipients were higher pp65 antigenemia than HLA-A * 2402 recipients (P = 0.007) and HLA-A * 1101 recipients (P = 0.028), HLA-A * 0201 recipients were higher pp65 antigenemia than HLA-A * 2402 (P = 0.02), the pp65 antigenemia showed no obvious difference among the rest of high-resolution HLA groups (P > 0.05).

CONCLUSIONHLA-DRB1 * 0901 alleles might be correlated with BMT recipients happened higher pp65 antigenemia, HLA-A * 2402 alleles might be correlated with BMT recipients happened lower pp65 antigenemia.

Adolescent ; Adult ; Bone Marrow Transplantation ; adverse effects ; Cytomegalovirus Infections ; immunology ; Female ; HLA Antigens ; genetics ; Humans ; Male ; Middle Aged ; Phosphoproteins ; blood ; Viral Matrix Proteins ; blood ; Viremia ; immunology

OBJECTIVETo determine the origin of 1 prenatally detected small supernumerary marker chromosome (sSMC) using SNP-chip technology, and to deduce the underlying mechanism.

METHODSThe fetal sample was subjected to karyotype analysis. The identified sSMC was subjected to genom wide scan using a SNP microarray chip. The results were validated with fluorescence in situ hybridization (FISH).

RESULTSThe karyotype of the fetus was determined as 46, X, +mar, which was verified by SNP microarray chip analysis as Yp11.2-11.3 duplication, along with loss of Yq11.2 region, FISH analysis has confirmed that the sSMC has derived from the Y chromosome.

CONCLUSIONThe karyotype of the fetus was determined as 46, X, idic(Y) (pter→ p11.2::11.2→ pter). Regional deletion of Yq11.2 has been associated with male azoospermia. SNP chip analysis can exclude minor deletions and duplications with a size of more than 1 Mb, which may be applied for verifying difficult cases as well as microdeletion and duplication syndromes upon prenatal diagnosis.

Adult ; Chromosome Disorders ; diagnosis ; embryology ; genetics ; Female ; Genetic Markers ; genetics ; Humans ; Karyotyping ; Male ; Oligonucleotide Array Sequence Analysis ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo determine the accuracy and clinical value of combining 64 multi-slice spiral computer tomography (MSCT) and serum amyloid A protein (SAA) in the preoperative staging of rectal cancer.

METHODSProspectively enrolled patients with rectal cancer from October 2007 to October 2008. The patients were randomly assigned into two groups: MSCT and SAA combined group: both MSCT and SAA combinative assessment were performed for preoperative evaluation; MSCT group: only MSCT was performed preoperatively for tumor staging. The accuracy of the preoperative T, N, M, and TNM staging and the concordance rate of predictive operative strategy were compared between the two groups.

RESULTSTotal of 225 cases with rectal cancer were enrolled in this study. There were 110 cases in MSCT and SAA combined group and 115 cases in MSCT group. The baseline characteristics was comparable between the two groups. For MSCT and SAA combined group, the accuracies of preoperative staging of T, N, M and TNM was 87.3%, 85.2%, 100% and 86.4%, respectively; and for MSCT group, the corresponding rates was 85.2%, 67.0%, 100% and 66.1%, respectively. Statistical differences was found in the accuracy of preoperative N and TNM staging between the two groups (P = 0.009 and 0.001, respectively). In addition, there was statistical difference in the accuracy of prediction to operative procedures between the two groups (94.7% vs. 81.7%, P = 0.003).

CONCLUSIONCombinative assessment of MSCT and SAA could improve the accuracy of preoperative staging, and thus provide higher predictive coincidence rate of operative procedures.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Preoperative Care ; Prospective Studies ; Rectal Neoplasms ; diagnosis ; surgery ; Serum Amyloid A Protein ; analysis ; Tomography, Spiral Computed ; methods