Using the concepts and methods of epigenetics and metabolomics, to investigate the overall action molecular mechanism of Chrysanthemi indici C (CIC), the anti-hepatitis B virus (HBV) active extracts from Flos chrysanthemi indici. The inhibitory effects of CIC on proliferation and hepatitis B surface antigen (HBsAg), hepatitis B envelope antigen (HBeAg) and HBV-DNA of HepG2.2.15 cells were detected by CCK-8 and antigen kit. The DNA methyltransferases (DNMTs)/ten-eleven-translocation-2 (TET2) equilibrium was detected by ELISA. Illumina 850K methylation chip, pyrosequencing and qPCR were used to determine the action pathway and target of CIC by GO and KEGG analysis. Cell metabolites were extracted with 80% methanol, and the changes of differential metabolites, differential metabolic pathways and cell microenvironment were detected by LC-MS and other metabolomics methods. The results showed that CIC could inhibit the proliferation, HBsAg, HBeAg and HBV-DNA of HepG2.2.15 cells obviously, down-regulate DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3a (DNMT3a) and DNA methyltransferase 3b (DNMT3b), up-regulate TET2, and restore the balance of DNMTs/TET2. The action targets of CIC were phospholipase C gamma 2 (PLCG2), phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3), 1-acylglycerol-3-phosphate O-acyltransferase 2 (AGPAT2), 5-hydroxytryptamine receptor 2B (HTR2B), nerve growth factor (NGF), mainly involved in lipid metabolism, inflammation mediated regulation of transient receptor potential (TRP), phospholipase D signaling and advanced glycation end product-receptor for AGE (AGE-RAGE) signaling in diabetic complications pathways. CIC could significantly affect fatty acid metabolism and had great influence on phenolic acid, alkaloid and lipid metabolites in cell microenvironment. These results suggest that the action mechanism of CIC may be the synergistic action of multiple pathways and multiple targets, including related inflammatory pathways, immune pathways and lipid metabolism, through regulating epigenetic expression balance and restoring the balance of cell microenvironment.

ObjectiveTo evaluate the curative effect difference between Tong's anterior mesh suspension and modified tension-free vaginal tape-obturator (TVT-O). Methods75 SUI cases were random divided into two groups: Group A (35 cases) were accepted Tong's anterior mesh suspension and group B (40cases) were accepted modified tension-free vaginal tape- obturator. Curative effects, operation safety,and complications and so on, were recorded and compared. ResultsThere had no statistical differences between these two groups on operation time, bleeding lose, operation injury, immediate postoperative urinary retention andcure rates, but the operation expense of group A was cheaper than group B [ (980. 74 ±212.45)yuan vs (2879.06 ±467. 13)yuan , P ＜0.05). ConclusionThe curative effect between two methods were similar. It's a little complicated and fit to be popularized in large hospital to modified tensionfree vaginal tape-obturator. Tong's anterior mesh suspension was performed under completely direct vision,not through pelvic cavity, and it was an economic, convenient and easy way, which not only fit to be popularized in basic hospital, but also a surgical remedial measure to the failures who accepted these operations,such as TVT, SPARC, IVS, MONARC, TOT, TVT-O, etc.

Objective To explore the relationship between methionine synthase reductase (MTRR) gene polymorphism and cerebral infarction in young and middle-aged patients. Methods The genotype of MTRR A66G was analyzed by polymerase chain reaction-restrictive fragment length polymorphism (PCR-RFLP) and the plasma homocysteine (Hcy) level was measured by high performance liquid chromatography in 105 young and middle-aged patients with cerebral infarction and 116 age-matched healthy controls. Results The genotype distribution and allele frequencies of MTRR A 66G gene between the 2 groups had no statistical significance (P＞0.05).Stratified analysis,performed according to whether cerebral infarction was complicated with hypertension,diabetes or coronary heart disease,indicated that the frequencies ofGG genotype and G allele in cerebral infarction patients without complications were obviously higher than those in controls (36.4％ vs.23.3％,62.1％ vs.52.2％),but no statistical significance was noted between them (P＞0.05).No statistical difference was observed between cerebral infarction patients with complications and controls (P＞0.05). The mean plasma Hcy level in patients and controls with GG genotype was significantly higher than that in patients and controls with AA genotype (P＜0.05). Conclusion No association between MTRR A 66G polymorphism and cerebral infarction is noted in young and middle-aged patients, while GG mutant homozygous ofMTRR A66G gene can significantly raise the plasma Hcy level.

INTRODUCTIONAntimicrobial stewardship programmes (ASP) can reduce antibiotic use but patient safety concerns exist. We evaluated the safety of prospective carbapenem review and feedback and its impact on carbapenem use and patient outcomes.

MATERIALS AND METHODSAfter 3 months implementation of our ASP, we compared patients with and without acceptance of ASP recommendations on the use of carbapenems. Primary outcome was 30-day mortality. Secondary outcomes included duration of carbapenem use, length of hospitalisation, clinical response, microbiological clearance, 30-day readmission and mortality at discharge.

RESULTSOf 226 recommendations for 183 patients, 59.3% was accepted. De-escalation, switching to oral antibiotics and antibiotic cessation comprised 72% of recommendations. Patients with acceptance of ASP recommendations had lower 30-day mortality and higher end-of-therapy clinical response despite shorter carbapenem duration (P <0.05). Predictors of 30-day mortality were Pitt bacteraemia score (adjusted odds ratio [aOR] 1.39, 95% confidence interval [CI], 1.11 to 1.74; P = 0.004) and non-acceptance of ASP recommendations (aOR 2.84, 95% CI, 1.21 to 6.64; P = 0.016).

CONCLUSIONOur prospective carbapenem review and feedback mainly comprising of reducing carbapenem use is safe.

Carbapenems ; therapeutic use ; Drug Utilization ; standards ; Feedback ; Guideline Adherence ; statistics & numerical data ; Humans ; Patient Safety ; Pharmaceutical Services ; Treatment Outcome

AIMTo study the effect of conjugated linoleic acid (CLA) on obese MSG mice with insulin resistance.

METHODSAbout four months old, obese MSG mice with insulin resistance were divided into control, CLA and rosiglitazone groups and drugs were administrated ig once a day. Body weights were recorded regularly, insulin and glucose tolerance were tested. In addition, serum insulin and TNF-alpha concentrations in serum and fat tissues were determined.

RESULTSCLA was shown to reduce the body weight and fat weight in MSG mice, but can not improve the abnormal insulin and glucose tolerance in these mice. Indeed, the serum insulin and TNF-alpha concentrations in the fat tissues of the group treated with CLA were higher than those in the models and the insulin sensitivity index was significantly lower than that in the model mice.

CONCLUSIONCLA can reduce the body weight of MSG mice, but can not improve the insulin resistance in these mice.

Adipose Tissue ; metabolism ; Animals ; Animals, Newborn ; Blood Glucose ; metabolism ; Body Weight ; drug effects ; Female ; Glucose Tolerance Test ; Insulin ; blood ; Insulin Resistance ; Linoleic Acids, Conjugated ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Obesity ; chemically induced ; metabolism ; Sodium Glutamate ; Tumor Necrosis Factor-alpha ; metabolism

The study was aimed to explore the mechanism of SYK and CBL family of ubiquitin ligases in Bufalin-induced HL-60 cells apoptosis. Cell viability was tested by trypan blue staining and apoptosis was detected by using flow cytometry. The expressions of CBL and CBL-b and the phosphorylation of SYK were detected by using immunoprecipitation and Western blot. The results showed that Bufalin inhibited HL-60 cell proliferation in time- and dose-dependent manners. IC(50) of suppressing cell viability at 24, 48 and 72 hours were about 26.3, 7.8 and 2.0 nmol/L respectively. The high dose of bufalin already induced apoptosis of HL-60 cells at 8 hours. SYK was quickly phosphorylated, and the expressions of CBL and CBL-b were down-regulated after treatment with Bufalin. It is concluded that SYK activation and CBL protein down-regulation may be involved in Bufalin-induced HL-60 cell apoptosis.
Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Protein-Tyrosine Kinases ; metabolism ; Proto-Oncogene Proteins c-cbl ; metabolism ; Signal Transduction ; Syk Kinase

Objective To design and clone all known and predicted coding genes of Epstein-Barr virus (EBV) as the cDNA probes for preparing the microarray for EBV detection, thereby to facilitate further investigation of the pathogenetic role of EBV. Methods Oligo 6.0 software, BLAST program and Primer Premier 5 software were employed to design and screen the cDNA probes of the whole EBV genome, whose length ranged from 300 to 600 mer each with high specificity. These cDNA probes obtained through PCR and reverse transcriptase (RT)-PCR amplification from the genomic DNA and RNA of B95-8 cells and nasopharyngeal carcinoma (NPC) tissue were cloned into T/A clone vector, followed by identification of these probes by sequencing analysis. Result and Conclusion A total of 85 gene fragments (BWRF1 genecontained 7 repeats of open reading frames) coding for proteins and 2 EBERs in EBV genome were successfully cloned, not including LF1 and LF3 genes that did not exist in EBV genome ofB95-8 cells, which provides the basis for preparing microarray to explore the role of EBV genome in its related diseases.

Objective To design and clone all known and predicted coding genes of Epstein-Barr virus (EBV) as the cDNA probes for preparing the microarray for EBV detection, thereby to facilitate further investigation of the pathogenetic role of EBV. Methods Oligo 6.0 software, BLAST program and Primer Premier 5 software were employed to design and screen the cDNA probes of the whole EBV genome, whose length ranged from 300 to 600 mer each with high specificity. These cDNA probes obtained through PCR and reverse transcriptase (RT)-PCR amplification from the genomic DNA and RNA of B95-8 cells and nasopharyngeal carcinoma (NPC) tissue were cloned into T/A clone vector, followed by identification of these probes by sequencing analysis. Result and Conclusion A total of 85 gene fragments (BWRF1 genecontained 7 repeats of open reading frames) coding for proteins and 2 EBERs in EBV genome were successfully cloned, not including LF1 and LF3 genes that did not exist in EBV genome ofB95-8 cells, which provides the basis for preparing microarray to explore the role of EBV genome in its related diseases.

OBJECTIVETo investigate the effect of HBP-A on meniscal injuries and the expressions of genes associated with pathological hypertrophy and calcification of the meniscusinduced by abnormal loading.

METHODSBovine meniscus explants were subjected to 25% strain at 0.3 Hz for 3 h and treated with 0.6 mg/mL of HBP-A. The cell viability in the meniscus explants after 72 hin culture was determined using live/dead staining and the expression levels of genes associated with pathological hypertrophy and calcification of the meniscus (ANKH, ENPP1, ALP, MMP13, and IL-1) were measured using real-time PCR and Western blotting. The conditioned medium was collected for testing sulfated glycosaminoglycan (GAG) release.

RESULTSThe number of dead cells, loss of proteoglycan content, and the expressions of ANKH, ENPP1, ALP and MMP13, and IL-1 at both the mRNA and protein levels were all significantly lower in the meniscus explants treated with 0.6 mg/mL HBP-A than in the explants with only 25% abnormal pressure stimulation (n=3, P<0.05).

CONCLUSIONHBP-A can effectively alleviate meniscal injuries induced by abnormal loading and suppress the expressions of genes related with pathological hypertrophy and calcification of the meniscus, and can serve as a potential drug for treatment of knee osteoarthritis.

Animals ; Calcinosis ; drug therapy ; Cattle ; Glucans ; pharmacology ; Hypertrophy ; Menisci, Tibial ; drug effects ; Osteoarthritis, Knee ; drug therapy ; Real-Time Polymerase Chain Reaction ; Tibial Meniscus Injuries ; drug therapy

Objective To clone and express VP_2 gene from HBoV,and the expressed VP_2 protein was as the antigen in order to detect serum from children in Wenling area with lower respiratory tract infections.Methods The VP_2 gene was reeombined with the genome of Baculovirus,which infected the insect cell.The fusion protein with HA tag was applied to confirm the specificity of expressed protein.Furthermore,the recombinant protein was observed using electron microscopy.The 176 serum from children in Wenling area with lower respiratory tract infections was screened using Western blot.Results The expressed VP_2 protein was more than 60% in total proteins from insect cell,and MWt about 60 ×10~3.The virus-like particle(VLP)Was observed using electron microscopy,and size about 20nm.The 176 serum from children in wenling area with lower respiratory tract infections was screened using Western blot.The HBoV positive rate Was 2.28%(4/176).Conclusion The VP_2 protein from human bocavims was expressed in insect cell successfully.Through HA tag the VP_2 protein wag specific,and then the assay using SDS-PAGE with Western blot could detect and screen the antibody in serum from children with lower respiratory tract infections rapidly and accurately.