Cytoskeletal Proteins ; biosynthesis ; genetics ; physiology ; Humans ; Neoplasm Metastasis ; Neoplasms ; pathology ; Signal Transduction

Animals ; Bone Neoplasms ; pathology ; Cell Adhesion ; Cell Culture Techniques ; methods ; Coculture Techniques ; Drug Screening Assays, Antitumor ; methods ; Humans ; Neoplasm Invasiveness ; Tumor Cells, Cultured

Objective The objective of this study was to investigate the levels of serum miR-21 and miR-34a in patients with advanced gastric cancer before and after Folfox chemotherapy and its relationship with chemotherapy efficacy.Methods Forty-five patients with advanced gastric cancer who underwent Folfox chemotherapy were enrolled from May 2015 to October 2015.The levels of miR-21 and miR-34a were analyzed by Real-time PCR before and after chemotherapy.The relationship between miR-21 and miR-34a levels and chemotherapy efficacy was analyzed.The predictive values of miR-21 and miR-34a on the efficacy of chemotherapy were evaluated by the receiver operating curve(ROC).Results After chemotherapy,6 cases(13.33%)were relieved,10 cases(22.22%)were stable disease(SD),29 cases(64.44%)were disease progress(PD),the objective response rate(ORR)was 13.33%,the disease control rate(DCR)was 35.56%,the median survival time was 8.1 months.The level of miR-21 was down-regulated and up-regulation for miR-34a after chemotherapy.They were statistically significant(P<0.05).The level of miR-21 in the chemotherapy group was lower than that in the ineffective group,and the level of miR-34a was higher than that of the ineffective group(P<0.05).MiR-21 was negatively correlated with the efficacy of chemotherapy(rs=-0.431,P<0.05),while miR-34a was positively correlated with it(rs=0.504,P<0.05).The median survival time was 7.6 months for low-level patients with miR-21 and 6.0 months for high-level patients.The median survival time of miR-34a patients was 5.5 months,compared with 8.3 months for high-level patients,with a statistically significant difference(P<0.05).The Log-Rank test showed that the median survival of patients with low miR-21 was higher than that of patients with high miR-21(P<0.05).The median survival of patients with high miR-34a was higher than that of patients with low miR-34a(P<0.05).The miR-21 combined with miR-34a in predicting chemotherapy efficacy under the curve(AUC)was 0.865,the sensitivity was 80.9% and the specificity was 89.8%.Conclusion miR-21 and miR-34a are correlated with chemotherapy efficacy median survival time of patients with advanced gastric cancer.

Objective Radiological grade of splenic injury was seldomly used in China trauma center now,though it had been established in 1994 by American Association for Surgery of Trauma (AAST) and widely used.The present study is aimed to analyze the imaging grade and clinical characteristics of traumatic splenic rupture in children,discuss the feasibility of conservative treatment,and the role of radiographic grading during clinical treatment.Methods Information (including age,gender,severity based on radiological findings,treatment strategies,and clinical outcome) regarding 59 hospitalized splenic injury patients whose injuries occurred between 2008 and 2014 was retrospectively analyzed.Results Between 2008 and 2014,59 pediatric patients with splenic injury were treated in our institution.Median age was 9.5 years (range,3 months to 16 years).Of all patients,41 (69.5%) were male.The injuries were primarily caused by traffic crash (45.7%),stumbling/falling from a height (38.9%).According to AAST,5 cases were grade Ⅰ,26 patients grade Ⅱ (44.1%),and 21 cases grade Ⅲ (35.6%),6 over grade Ⅳ,and only one was unclear.Of all patients,25 cases were with the other organs complications.All patients underwent fasting,bed rest,and antibiotics.Only 1 case was transferred to operation during the conservative treatment.Forty-nine patients underwent with CT scan over 2 times.Conclusion Imaging classification helps guide clinical treatment.Conservative treatment is feasible for traumatic splenic injury in children.Early imaging classification of splenic injury may be helpful in clinical judgment,and reduce children radiation exposure.

Objective:To screen for the pathogenesis-related genes of osteosarcoma and to assess their roles for the de- velopment of osteosareoma.Methods:Total RNA was extracted from 3 ATCC osteosarcoma cell lines and an osteoblastic cell line and was used to synthesize biotinylated cRNAs;the latter were hybridized to Affymetrix~(?)GeneChip~(?)U133A ar- rays and a gene with more than 2 folds of change was selected.Ten of the differentially expressed genes were chosen and the primers were designed and the synthesized.Then SYBR~(?)Green real-time PCR(RT-PCR)method was used to detect the expression of the 10 genes in 9 fresh osteosarcoma specimens.ABI Prism 7 000 system was used to analyze the differ- ent expression between osteosarcoma cell line and osteoblastic cell line.Results:We identified 58 up-regulated and 142 down-regulated genes in the 3 osteosareoma cell lines.Many of the genes were firstly reported to be related to the patho- genesis of osteosarcoma.These differentially expressed genes were mainly involved in energy and material metabolism,on- cogene,signal transduction gene,transcription- related genes,cell cycle-related genes,cell apoptosis-related gene,im- mune response gene,tumor suppressor genes,etc.The array results of 10 randomly selected genes were further verified by the RT-PCR in 9 fresh osteosarcoma specimens.Conclusion:Many genes are involved in the pathogenesis of osteosarcoma. Gene microarray can help to discover the genes related to the pathogenesis of osteosarcoma,which may lay a foundation for studying the molecular mechanism of osteosarcom.

OBJECTIVETo evaluate the effect and safety of L-carnitine in the treatment of idiopathic oligoasthenozoospermia based on current clinical evidence.

METHODSWe searched the Cochrane Library, PubMed, MEDLINE, EMBASE, CNKI, VIP, CBM and Wanfang Database from the establishment to April 2014 for the published literature on the treatment of idiopathic oligoasthenozoospermia with L-carnitine. We conducted literature screening, data extraction, and assessment of the methodological quality of the included trials according to the inclusion and exclusion criteria, followed by statistical analysis with the RevMan 5. 2 software.

RESULTSSeven randomized controlled trials involving 751 patients with idiopathic oligoasthenozoospermia met the inclusion criteria, and 678 of them were included in the meta-analysis. L-carnitine treatment achieved a significantly increased rate of spontaneous pregnancy as compared with the control group (RR = 3.2, 95% CI 1.74 to 5.87, P = 0.0002). After 12-16 and 24-26 weeks of medication, total sperm motility (WMD = 5.21, 95% CI 2.78 to 7.64, P < 0.0001 and WMD = 9.29, 95% CI 1.28 to 17.29, P = 0.02) and the percentage of progressively motile sperm (WMD = 12.44, 95% CI 4.58 to 20.31, P = 0.002 and WMD = 9.76, 95% CI 3.56 to 15.97, P = 0.002) were remarkably higher than those in the control group, but no statistically significant differences were observed in sperm concentration between the two groups (WMD = 4.91, 95% CI -2.63 to 12.45, P = 0.2 and WMD = 0.93, 95% CI -3.48 to 5.34, P = 0.68). After 12-16 weeks of treatment, the percentage of morphologically abnormal sperm was markedly decreased in the L-carnitine group as compared with the control (WMD = -2.48, 95% CI -4.35 to -0.61, P = 0.009), but showed no significant difference from the latter group after 24-26 weeks (WMD = -4.38, 95% CI -9.66 to 0.89, P = 0.1). No statistically significant difference was found in the semen volume between the two groups after 12-16 or 24-26 weeks of medication (WMD = -0.13, 95% CI -0.43 to 0.18, P = 0.42 and WMD = 0.28, 95% CI -0.02 to 0.58, P = 0.07). No serious L-carnitine-related adverse events were reported in 4 of the randomniized controlled trials.

CONCLUSIONThe current evidence indicates that L-carnitine can improve spontaneous pregnancy and semen parameters in the treatment of idiopathic oligoasthenozoospermia, with no serious adverse reactions.

Asthenozoospermia ; drug therapy ; Carnitine ; adverse effects ; pharmacology ; Female ; Humans ; Male ; Pregnancy ; Pregnancy Rate ; Randomized Controlled Trials as Topic ; Semen Analysis ; Sperm Count ; Sperm Motility

The effects of dietary supplementation with Clostridium butyricum on growth performance and humoral immune response in Miichthys miiuy were evaluated. One hundred and fifty Miichthys miiuy weighing approximately 200-260 g were divided into five groups and reared in 15 tanks with closed circuiting culture system. The animals were fed 5 diets: basal diet only (control) or supplemented of the basal diet with C. butyricum at doses of 10(3) (CB1), 10(5) (CB2), 10(7) (CB3) or 10(9) (CB4) CFU/g. Compared with the control, the serum phenoloxidase activity was significantly increased by the supplementation (P<0.05), acid phosphatases activity was increased significantly (P<0.05) at the doses of 10(9) CFU/g. Serum lysozyme activity peaked at dose of 10(7) CFU/g and in the skin mucus at dose of 10(9) CFU/g. Immunoglobulin M level in the serum and skin mucus was increased except at dose of 10(3) CFU/g (P<0.05). The growth at the dose of 10(9) CFU/g was higher than that of the control (P<0.05). It is concluded that supplementation of C. butyricum can mediate the humoral immune responses and improve the growth performance in Miichthys miiuy.
Animal Feed ; microbiology ; Animals ; Antibody Formation ; physiology ; Clostridium butyricum ; immunology ; Dietary Supplements ; microbiology ; Fishes ; growth & development ; immunology ; Probiotics ; administration & dosage

NS2 is a nonstructural protein of Periplaneta fuliginosa densovirus(PfDNV) with a molecular mass of 30 kD,whose function is not yet clearly understood. In order to study the expression,subcellular distribution and the function of NS2 protein,the coding region of NS2 was amplified from the hindgut tissue of cockroaches infected with PfDNV by RT-PCR and then the recombinant prokaryotic expression vector pET28a-NS2 was constructed. The recombinant plasmid was transformed into E. coli BL21(DE3) to express the 6?His fusion protein in the bacteria. After purification,the fusion protein was injected into New Zealand rabbits to prepare polyclonal antibody. The specificity of the anti-NS2 antibody was successfullyproved by western blotting on the eukaryotic expressed products of NS2 protein.Meanwhile,the full sequence of ns2 gene was also cloned into the eukaryotic expression vector pAC. The recombinant plasmid pAC-NS2 was then transfected into Schneider line 2(S2) cells to express NS2 protein in the insect cells. The subcellular localization of NS2 in the insect cells was then investigated by indirect immunofluorescence technique using the anti-NS2 polyclonal antiserum. The confocal laser scanning microscope observation showed that NS2 protein was located primarily in the cytoplasm with some punctate nuclear staining.

Objective To investigate the effects of total flavonoids of litchi chinensis sonn (TFL) on cell proliferation and the molecular mechanism in rat hepatic stellate cells (HSC-T6) activated by growth factor-β1 (TGF-β1). Methods HSC-T6 cells were treated by 0.25%Trypsin-EDTA and then were digested into single cell suspension by DMEM (10%FBS included), which were mixed with TGF-β1 (5μg/L). (1) MTT method was used to detect the proliferation of HSC-T6 cells. Cells were cultured in 96-well plate and were treated by different concentrations of TFL including TGF-β1 group, the control group (5‰DMSO included), and different concentrations of TFL groups (80, 160, 320, 640 and 800 mg/L TFL). Each group has three wells. The absorbance (A) value was measured by enzyme standard meter at the 490 nm wavelength after 24 h, 48 h and 72 h treatment. The cell inhibitory rate was calculated. The subsequent experimental drug concentration and drug treatment time were determined according to half inhibitory concentration (IC50). (2) The expression levels of NF-κB andα-SMA mRNA were detected by PCR (for mRNA) and Western blot assay (for protein). Cells were cultured in the 10 cm culture dish and were divided into different TGF-β1 groups, including TGF-β1 group, the control group (5‰DMSO included), and different concentrations of TFL groups (125, 250 and 500 mg/L TFL). After 48 h, related indicators were measured. Results At the same treatment time point, with the increased concentrations of TFL, A values were gradually decreased, and the cell inhibitory rates were gradually increased. There were no significant differences in the expressions of NF-κB andα-SMA mRNA between TGF-β1 group and control group. And there were no significant differences in the expressions of NF-κB andα-SMA mRNA between TFL125 group, TGF-β1 group and control group. There was a gradually decrease in the expressions of NF-κB andα-SMA mRNA and protein with the increased concentrations of TFL. Conclusion TFL can inhibit TGF-β1-induced HSC-T6 cell proliferation, which is involved in the inhibited expressions of NF-κB andα-SMA to anti-fibrotic effects in liver fibrosis.

Objective To investigate the role of TNSALP gene detection in prenatal diagnosis of HPP. Method The clinical data and the results of complete exon sequencing of TNSALP gene in one neonate with low alkaline phosphatase (HPP) were analyzed retrospectively. Peripheral bloods from his family members were collected. The amniotic fluid cell in fetuses at 17 weeks was tested for candidate gene mutations by Sanger sequencing. Results Mainly manifestations in 6-day-old baby were multiple fractures, limb shortening and bending and dyspnea. He died of respiratory failure 9 days after birth. The serum alkaline phosphatase was decreased and serum calcium was decreased slightly; serum phosphorus, serum 25 hydroxyvitamin-D and parathyroid hormone were normal. X-ray showed that the whole body bone was very poorly mineralized, and the long diaphysis was enlarged with shape of a cup at the end and multiple fractures existed. Gene sequencing revealed a complex heterozygous missense mutation in the TNSALP gene, including the heterozygous missense mutation c.542C>T in exon sixth causing 181st amino acids changed from serine to leucine (p.S181L), and tenth exon heterozygous missense mutation in c.1016G>A causing 339th amino acid changed from glycine to glutamic acid (p.G339E). The parental phenotypes were normal. The c.542C>T mutation is inherited from his father and the c.1016G>A mutation is inherited from his mother. These two mutations were not detected in the fetus. Conclusion TNSALP gene analysis can be applied to the diagnosis and prenatal diagnosis of HPP.