Objective To explore the effect and molecular mechanism of HDAC inhibitor SNDX-275 inhibiting cell proliferation in ErbB2-overexpressing breast cancer BT474 cells.Methods Breast cancer BT474 cells were treated with HDAC inhibitor SNDX-275,setting as test group,and the cell line treated with phosphate buffered saline (PBS)as control.The concentration of SNDX-275 were 0,0.5,1 .0,2.0,3.0,4.0 μmol/L respectively.Cell proliferation was analyzed by MTS assay and colony formation assay,the expressions of ErbB2, ErbB3,p-Akt were analyzed by Western blotting,and the expressions of miR-125a,miR-125b were analyzed by RT-PCR.After transfecting miRNA125 inhibitor into BT474 cells,the inhibition rate of SNDX-275 was tested by MTS assay .Results MTS result showed that SNDX-275 inhibited cell proliferation in BT474 cells in a dose-dependent manner.The inhibition rate of 4.0 μmol/L SNDX-275 was about (68.00 ±4.45)%.Clone assay indicated SNDX-275 could inhibit the proliferation of BT474 cells.Western blotting result indicated that SNDX-275 significantly inhibited the protein expressions of ErbB2,ErbB3 and p-Akt,RT-PCR result illustrated 2 μmol/L SNDX-275 could increase the expressions of miR-125a and miR-125b about 3.22 ±1 .17,5.42 ±0.38 times compared with the PBS control respectively,the difference has a statistical significance (t =4.338,P =0.049;t =21 .805,P =0.002).MTS result indicated that compared with the PBS control,the inhibition rate of SNDX-275 group was (56.97 ±3.56)%,while the inhibition rate of SNDX-275 and miRNA125 inhibitor group
was (10.67 ±2.21 )%,with a statistical significance(t =-10.993,P =0.008).Conclusion SNDX-275 could inhibit cell proliferation of ErbB2-overexpressing breast cancer BT474 cells,by inhibiting ErbB2-ErbB3-Akt sig-nal pathway through up-regulating miR-125a and miR-125b.

Sugarcane bagasse modified by polyethylenimine (PEI) and glutaraldehyde (GA) was used as a carrier to immobilize Clostridium acetobutylicum XY16 in the process of butanol production. The effects of chemically modified sugarcane bagasse on batch and repeat-batch fermentations were investigated. Batch fermentation was conducted with an addition of 10 g/L modified sugarcane bagasse and 60 g/L glucose, resulting in a high solvent concentration of 21.67 g/L and productivity of 0.60 g/(L x h) with the treatment of 4 g/L PEI and 1 g/L GA. Compared to the fermentations by free cells and immobilized cells on unmodified sugarcane bagasse, the productivity increased 130.8% and 66.7%, respectively. The fibrous-bed bioreactor also maintained a stable butanol production during repeat-batch fermentations, achieving a maximum productivity of 0.83 g/(L x h) with a high yield of 0.42 g/g.
Batch Cell Culture Techniques ; Bioreactors ; Butanols ; metabolism ; Cells, Immobilized ; Cellulose ; metabolism ; Clostridium acetobutylicum ; metabolism ; Fermentation ; Saccharum ; chemistry

Aged ; Branchioma ; pathology ; Carcinoma, Squamous Cell ; pathology ; Head and Neck Neoplasms ; pathology ; Humans ; Male

Aim To investigate the effect of asiatic acid on apoptosis and autophagy in human glioblastoma T98G cells. Methods MTT colorimetry was employed to assay the cellular proliferating activity. The fluores-cence microscope and Hoechst 33258 staining were used to detect the morphological changes. The cell ap-optosis and autophagy were analyzed by flow cytometry with Annexin-V/7-AAD and MDC staining respective-ly. The expressions of associated proteins were detected by Western blot to analyze the mechanism of apoptosis and autophagy. Results MTT assay showed that the growth of T 9 8 G cells was inhibited by asiatic acid ( IC50 =46. 3 μmol · L-1 ) . Annexin V/7-AAD stai-ning and Western blot revealed that asiatic acid in-duced apoptosis in T98 G cells by reducing the expres-sion of Akt, decreasing the mitochondrial membrane potential, and increasing the expression of Caspase-3. MDC staining and Western blot showed that the per-centage of MDC-positive cells was decreased and the expressions of Beclin-1 , LC3-II and Atgs were inhibi-ted by asiatic acid treatment. 5 μmol·L-1 chloroquine was used to up-regulate the expressions of LC3-Ⅱand Beclin-1 . Asiatic acid-inhibited autophagy was blocked and the total apoptotic rate was reduced remarkably. Conclusion Asiatic acid suppresses T98 G cells pro-liferation by inducing apoptosis and inhibiting cell au-tophagy, and the very role of inhibiting autophagy could promote apoptosis to a certain extent.

OBJECTIVETo observe the effect of salidroside on tic behavior and in vivo dopamine DA) and serotonin (5-HT) levels in Tourette syndrome (TS) model rats.

METHODSForty rats were randomly divided into the blank control group, the TS model group, the haloperidol-treated group (0.5 mg/kg x d(-1)), and the salidroside-treated group (50 mg/kg x d(-1)), 10 in each group. TS rat model was induced by imino-dipropio-nitrile (IDPN). Peritoneal injection of haloperidol and salidroside was started from the 4th day of modeling in the haloperidol-treated group and the salidroside-treated group respectively. Normal saline was peritoneally injected to rats in the blank control group and the TS model group respectively. Stereotyped behavior was scored, and changes of DA and 5-HT levels in blood and striatum were measured before modeling, after modeling, and after intervention.

RESULTSCompared with the blank control group, the score of the tic behavior was elevated (P < 0.01) , levels of DA and 5-HT in plasma and striatum were reduced in the model group (P < 0.01, P < 0.05). Compared with the same group after modeling, the tic behavior score decreased and plasma DA levels increased in the two treated groups after intervention (P < 0.01). 5-HT content increased in the salidroside-treated group (P < 0.01). Compared with the model group after intervention, the tic behavior score was significantly reduced (P < 0.01), and DA levels in plasma and striatum were elevated (P < 0.01, P < 0.05) in the salidroside-treated group and the haloperidol-treated group. Compared with the haloperidol-treated group, the tic behavior score increased (P < 0.01), DA levels in plasma and striatum were lowered (P < 0.01, P < 0.05), the 5-HT level increased in plasma and striatum (P < 0.01, P < 0.05) in the salidroside-treated group.

CONCLUSIONSIn the salidroside-treated group, the tic behavior was significantly reduced, and DA levels in plasma and striatum were elevated. Its mechanism might be related to regulating activities of dopamine neurons in striatum.

Animals ; Corpus Striatum ; Dopamine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Glucosides ; pharmacology ; therapeutic use ; Haloperidol ; Phenols ; pharmacology ; therapeutic use ; Rats ; Serotonin ; Stereotyped Behavior ; Tics ; drug therapy ; Tourette Syndrome ; drug therapy

OBJECTIVETo determine the relationship of serum hepatocyte growth factor (HGF) and matrix metalloproteinase-9 (MMP-9) levels with the disease activity of systemic lupus erythematosus (SLE).

METHODSSerum levels of HGF and MMP-9 were measured by ELISA in 36 patients with SLE and 30 healthy subjects as controls.

RESULT(1)Significantly increased serum level of HGF was found in SLE patients as compared with that in healthy controls (P<0.001), but serum level of MMP-9 in SLE patients decreased (P<0.001). Serum level of HGF was significantly decreased after treatment in SLE patients (P<0.05), but serum level of MMP-9 was increased (P<0.05). (2)Serum level of HGF was markedly higher in patients with active disease (24 cases) than those with inactive disease (P<0.05), but serum level of MMP-9 was lower (P<0.05). (3)Significantly increased serum level of HGF was found in patients with renal damage (16 cases) than those without (P<0.001), but serum level of MMP-9 was lower in patients with renal damage than those without (P<0.005). (4)Serum level of HGF was higher in patients with arthritis (23 cases) than those without (P<0.01), but serum level of MMP-9 had no significant difference in two groups (P>0.05). (5)The area of ROC curve was 0.707 and the sensitivity was 66.7% if serum level of HGF was served as diagnostic standard. The area of ROC curve was 0.984 and the sensitivity was 97.2% if serum level of MMP-9 was served as diagnostic standard. The sensitivity was 66.7% (24/36) if the two markers were examined simultaneously.

CONCLUSIONThe data suggest that HGF and MMP-9 could be involved in the pathogenesis of SLE, and serum levels of HGF and MMP-9 be used as markers to monitor disease activity, renal damage, disease progression and amelioration in SLE.

Adolescent ; Adult ; Female ; Hepatocyte Growth Factor ; blood ; Humans ; Lupus Erythematosus, Systemic ; blood ; Male ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; ROC Curve

OBJECTIVETo explore the value of employing the small intestinal feeding tube in treating high position intestinal obstruction of newborn infant.

METHODFive newborn infants (3 males and 2 females; 1 premature infant and 4 fully-mature infants; 2 had membranous atresia of duodenum, 1 had annular pancreas, and 2 had proximal small intestine atresia; 1 infant had malrotation). The duodenal membrane-like atresia and the blind-end of small intestine were removed and intestinal anastomosis was performed, which was combined with intestinal malrotation removal. Before the intestinal anastomosis surgery, the anesthetist inserted via nose a 6Fr small intestinal ED tube, made by CREATE MEDIC CO LTD of Japan[

REGISTRATION NUMBERthe State Food and Drug Administration-instrument (Im.) 2007-NO.2661620]. Twenty-four hours after surgery, abdominal X-ray plain film was taken and patients were fed with syrup; 48 hours later, formula milk was pumped or lactose-free milk amino acids were given by intravenous injection pump through the feeding tube. The amount of milk and fluids was gradually increased to normal amount according to the condition. In initial 3 days the intravenous nutrition was given and one week after operation, the infants were fed through mouth in addition to pumping milk through the tube and stopped infusion. Ten to 22 days after operation, the tube was removed and the infant patients were discharged.

RESULTAll the five infants showed that the feeding through the nutrition tube was accomplished and the time of venous nutrition was reduced and fistula operation was avoided. None of the infants on question was off the tube and no jaundice exacerbation was found and the liver function was also found normal. At the very beginning, the tube was occasionally blocked by milk vale in one infant and after 0.9% sodium chloride solution flushing patency restored. After that, the feeding tube was washed once with warm water after feeding. In one infant vomiting occurred due to enough oral milk. The photograph of upper gastrointestine did not show anastomomotic stricture or fistula, or intestinal obstruction. After pulling out the tube, the symptoms disappeared and then the patient was discharged. One child was found to have diarrhea with no lactose nutrition liquid and given compound lactic bacteria preparations for oral administration, the symptom disappeared. In the 5 cases, the shortest hospital stay was 10 days and the longest was 22 days, the average stay was 16 days. Three to 5 days after operation the weight restored to birth weight, the weight had increased, when discharged, to an average of 5.5 g (kg·d).

CONCLUSIONThe small intestinal feeding tube was very effective for the postoperative nutrition maintenance of high position intestinal obstruction in newborn infants.

Anastomosis, Surgical ; Enteral Nutrition ; instrumentation ; methods ; Female ; Humans ; Infant, Newborn ; Intestinal Atresia ; surgery ; Intestinal Obstruction ; surgery ; Intestine, Small ; abnormalities ; surgery ; Intubation, Gastrointestinal ; instrumentation ; methods ; Length of Stay ; Male ; Nose ; Postoperative Care ; methods ; Retrospective Studies ; Time Factors ; Weight Gain

AIMTo identify and characterize a novel gene with potential roles in testis development and spermatogenesis.

METHODSA cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with probes of human or mouse adult and fetal testes. Differentially expressed genes were isolated and sequenced. RT-PCR was used to test the tissue distribution of the genes of interest and in situ hybridization was performed to localize the gene expression in the mouse testis. A range of bioinformatical programs including Gene Runner, SMART, NCBI Blast and Emboss CpGPlot were used to characterize the new gene's feature.

RESULTSA novel testis-specific gene, NYD-SP5, was differentially expressed in fetal and adult testes. The deduced protein structure of NYD-SP5 was found to contain an IQ motif (a short calmodulin-binding motif containing conserved Ile and Gln residues), a Carbamate kinase-like domain, a Zn-dependent exopeptidase domain and a lactate dehydrogenase (LDH) C-terminal-like domain. RT-PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues examined. In situ hybridization and RT-PCR examinations revealed that the expression of NYD-SP5 was confined in the male germ cell but not present in the somatic cell in the testes.

CONCLUSIONNYD-SP5 is a newly found testis-specific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme.

Amino Acid Sequence ; Animals ; Base Sequence ; DNA, Complementary ; In Situ Hybridization ; Male ; Mice ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Proteins ; chemistry ; genetics ; physiology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid ; Spermatogenesis ; genetics ; Testis ; growth & development ; metabolism

AIMTo identify the genes specifically expressed in human adult and fetal testes and spermatozoa.

METHODSA human testis cDNA microarray was established. Then mRNAs of human adult and fetal testis and spermatozoa were purified and probes were prepared by a reverse transcription reaction with mRNA as the template. The microarray was hybridized with probes of adult and fetal testes and spermatozoa. The nucleic acid sequences of differentially expressed genes were determined and homologies were searched in the databases of GenBank.

RESULTSA novel human testis-specific gene, PKH-T, was identified by hybridizing adult and fetal testis and spermatozoa probes with a human testis cDNA microarray. The cDNA of PKH-T was 1 069 bp in length. The cDNA sequence of this clone was deposited in the Genbank (AY303972) and PKH-T was also determined as Interim GenSymbol (Unigene, HS.38041). PKH-T contained most PKH conserved motif. The 239 amino acid sequences deduced from the 719 bp open reading frame (ORF) had a homology with the gene PKH (U89606). PKH-T was specifically and strongly expressed in the testis. Comparison of the differential expressions of PKH and PKH-T in testes of different developmental stages indicated that PKH-T was expressed in the adult testis and spermatozoa, while PKH, in the adult, fetal and aged testes. PKH-T had no expression in the testis of Sertoli cell only and partially spermatogenic arrest patients.

CONCLUSIONPKH-T is a gene highly expressed in adult human testis and spermatozoa. It may play an important role in spermatogenesis and could be related to male infertility.

Adult ; Amino Acid Sequence ; DNA, Complementary ; biosynthesis ; genetics ; Female ; Gene Expression Regulation, Enzymologic ; genetics ; Humans ; Infertility, Male ; genetics ; Isoenzymes ; biosynthesis ; genetics ; Male ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Pregnancy ; Pyridoxal Kinase ; biosynthesis ; genetics ; RNA Splicing ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sertoli Cells ; metabolism ; Spermatogenesis ; genetics ; Spermatozoa ; metabolism ; Testis ; embryology ; enzymology ; growth & development ; Tissue Distribution

OBJECTIVESTo investigate the relationship between chromosome balanced translocation t(1;12) (q24;q24) and spermatogenesis in infertile twin brothers.

METHODSFor twin brothers, karotype were determined. The levels of testosterone, FSH and LH were detected. YRRM1, DAZ and DYS240 were analyzed. In younger brother a biopsy was taken from testis.

RESULTSChromosome analysis for both twin brothers revealed a karotype of 46, XY, t(1;12) (q24;q24). Sperm count was less than 1.0 x 10(6)/ml. There was no deletion for YRRM1, DAZ and DYS240 gene on Y chromosome. Photomicrograph of seminiferous tubules showed the arrest of spermatogenesis.

CONCLUSIONSChromosome balanced translocation t(1;12) (q24;q24) may be the cause of the spermatogenesis arrest in infertile twin brothers. Gene in the point of translocation may be related to spermatogenesis.

Adult ; Biopsy ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 12 ; Diseases in Twins ; genetics ; Humans ; Infertility, Male ; genetics ; Male ; Spermatogenesis ; genetics ; Testis ; pathology ; Translocation, Genetic ; genetics