Objective To explore the effects of the autoantibody against the second extracellular loop of ?3-adrenoceptor(?3AR)from children with heart failure on isolated cardiomyocytes of adult rats.Methods The synthetic peptide corresponding to the sequence of the se-cond extracellular loop(176-202 amino acid residues)of the human ?3AR was used as the antigen to screen the anti-?3AR autoantibodies from the sera of the 50 children with heart failure and 50 healthy children by enzyme linked immunosorbent assay(ELISA);IgG in the children with heart failure of positive autoantibodies sera was purified.The effects of purified IgG per each group on contractile response of isolated cardiomyocytes of adult rats were observed using motion-edge detection system and double-provocation fluorescence photomultiplier system.Results Among the 50 children with heart failure,the anti-?3AR autoantibodies positive rate was 30%(15/50 cases),and ob-viously higher than that of healthy children[12%(6/50)cases,P

OBJECTIVE To investigate the genotypes of plasmid-mediated AmpC beta-lactamases carried in Klebsiella pneumoniae,to provide reasonable use of antibacterial agents and to reduce the spread of these drug resistant genes.METHODS Two hundred and eighteen strains of K.pneumoniae were performed in Tongji Hospital in Wuhan.Plasmid-mediated AmpC beta-lactamases-producing strains were screened by improved three-dimensional method and identified by multiplex PCR.DNA sequencing method was used to confirm these drug resistant strains.The MIC of 15 kinds of antibacterial agents against the clinical isolates was detected by double agar dilution method.RESULTS Thirteen strains of K.pneumoniae were detected by improved three-dimensional method.The detection rates were 5.96%.There were seven of thirteen positive strains of K.pneumoniae through improved three-dimensional method harboring DHA-1 type plasmid-mediated AmpC beta-lactamase by multiplex PCR and confirmed by DNA sequencing.Plasmid-mediated AmpC beta-lactamase producers revealed a highly drug resistance to cephalosporins,monobactams,beta-lactam/beta-lactamase inhibitor combinations,aminoglycosides and fluoroquinolones.Only imipenem was susceptible to all of these detected strains.CONCLUSIONS DHA-1 plasmid-mediated AmpC beta-lactamase is detected in K.pneumoniae strains from Tongji Hospital.The detection rate is 3.2%.The pAmpC-producing strains reveal multi-drug resistance.Only imipenem is susceptible to them.

OBJECTIVETo investigate the chemical constituents of the dried sorophore of cultured Cordyceps militaris.

METHODCompounds were isolated and purified by macroporous adsorption resin and silica gel column chromatography. Their chemical structures were elucidated on the basis of physicochemical properties and spectral data (IR, FAB-MS, 1H-NMR and 13C-NMR).

RESULTNine compounds were isolated and identified as: ergosta-4, 6, 8 (14)-tetraen-3-one (1), citrostadienol (2), tetracosanoic acid 2, 3-dihydroxypropyl ester (3), ergosterol (4), ergosterol peroxide (5), ergosta-7, 22-dien-3beta, 5alpha, 6beta-triol (6), cordycepin (7), adenosine (8), N-(2-hydroxyethyl) adenosine (9), respectively.

CONCLUSIONCompounds 1-3, 6, 9 were separated from the sorophore of cultured C. militaris for the first time.

Cordyceps ; chemistry ; Culture Techniques ; Fruiting Bodies, Fungal ; chemistry

Objective To investigate the relationship between the promoter of IL-12B gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without Helicobacter pylori(Hp) infection in children and adolescent.Methods Mucosal biopsies were obtained from 132 children and adolescent (patient group),including 100 children with chronic gastritis and 32 children with duodenal ulcer,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin (HE),and gastritis was graded according to the Sydney system.Serology,urease test and histology were taken to assess Hp status.Genomic DNA was obtained from peripheral blood or gastric biopsies of patients and 102 healthy children as normal control group.The promoter of IL-12B +1188A/G gene polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing.The genotype distributions and allele frequencies were compared between the study group and the normal control group,and the association of genotypes with clinicopathological features was studied.IL-12B mRNA level expressions in gastric mucosa were confirmed by reverse transcription PCR biopsy-based tests.Results The genotype distributions and allele frequencies of IL-12B +1188A/G gene polymorphisms were similar in gastric upper gastrointestinal diseases and healthy subjects.The IL-12B +1188A/G gene polymorphisms were not associated with Hp status.IL-12B+1188A/G gene polymorphisms did not affect IL-12B mRNA level expressions and were not associated with the degree of antrum chronic inflammation.Conclusions These data suggest that IL-12B+1188A/G gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.

OBJECTIVETo investigate the feasibility and short-term outcomes of total laparoscopic sigmoid and rectal surgery combined with transanal endoscopic microsurgery(TEM).

METHODSThe clinical data of 26 patients with colorectal carcinoma treated by total laparoscopic surgery with TEM between May 2010 and May 2011 in the Shanghai Ruijin Hospital were retrospectively analyzed.

RESULTSAll the 26 operations were successfully accomplished laparoscopically. There was no conversion to open procedure. No diverting ileostomy was made. The mean operative time was (151.6±25.9) min. The mean blood loss was (200.2±114.7) ml. The mean time to first flatus was (2.0±0.5) d. The mean tumor size was (3.0±0.7) cm and all resection margins were negative. The mean number of lymph nodes harvested was (12.9±2.2). Six patients developed postoperative anastomotic leakage, all of who had tumors in the lower rectum. There were no ureteral injury, intestinal obstruction, or pulmonary infection.

CONCLUSIONSTotal laparoscopic sigmoid and rectal surgery combined with TEM is a safe and feasible minimally invasive surgery. It is an improvement by combining laparoscopic skills with the concept of natural orifice transluminal endoscopic surgery.

Adult ; Aged ; Anal Canal ; surgery ; Colorectal Neoplasms ; surgery ; Endoscopy, Gastrointestinal ; methods ; Female ; Humans ; Laparoscopy ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the properties of screw-bone interface of expansive pedicle screw (EPS) in osteoporotic sheep by micro-CT and histological observation.

METHODSSix female sheep with bilateral ovariectomy-induced osteoporosis were employed in this experiment. After EPS insertion in each femoral condyle, the sheep were randomly divided into two groups: 3 sheep were bred for 3 months (Group A), while the other 3 were bred for 6 months (Group B). After the animals being killed, the femoral condyles with EPS were obtained, which were three-dimensionally-imaged and reconstructed by micro-CT. Histological evaluation was made thereafter.

RESULTSThe trabecular microstructure was denser at the screw-bone interface than in the distant parts in expansive section, especially within the spiral marking. In the non-expansive section, however, there was no significant difference between the interface and the distant parts. The regions of interest (ROI) adjacent to EPS were reconstructed and analyzed by micro-CT with the same thresholds. The three-dimensional (3-D) parameters, including tissue mineral density (TMD), bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), were significantly better in expansive sections than non-expansive sections (P less than 0.05). Histologically, newly-formed bony trabeculae crawled along the expansive fissures and into the center of EPS. The newly-formed bones, as well as the bones at the bone-screw interface, closely contacted with the EPS and constructed four compartments.

CONCLUSIONSThe findings of the current study, based on micro-CT and histological evaluation, suggest that EPS can significantly provide stabilization in osteoporotic cancellous bones.

Animals ; Bone Screws ; Bone and Bones ; diagnostic imaging ; pathology ; Female ; Osteoporosis ; diagnostic imaging ; pathology ; Sheep ; Tomography, X-Ray Computed

Objective: To observe the effect of miR-155 on angiotensin Ⅱ (AngⅡ)-induced mice vascular smooth muscle cell (VSMC) phenotype switching with its possible mechanism. Methods: Primary cultured mice VSMCs were treated by AngⅡ at different concentrations and time periods, relevant expressions of miR-155 were examined by RT-PCR. qRT-PCR was conducted to determine miR-155 changes in Blank control group, miR-155 mimics group, miR-155 mimics negative control (NC) group, miR-155 inhibitor group and miR-155 inhibitor NC group. Western blot analysis was performed to measure the effect of miR-155 on AngⅡ-enforced ERK1/2 and mTOR signaling pathway in Blank control group, AngⅡ group, miR-155 mimics group, AngⅡ+miR-155 mimics group, miR-155 inhibitor group and AngⅡ+miR-155 inhibitor group; to detect the impact of miR-155, rapamycin (Rap) and U0126 on AngⅡ promoted VSMC phenotype switching in Blank control group, AngⅡ group, miR-155 mimics group, AngⅡ+miR-155 mimics group, AngⅡ+U0126 group and AngⅡ+Rap group, and to detect protein expressions of SM22α, α-SM-actin (contractile phenotype marker protein) and OPN (synthetic phenotype marker). Results: AngⅡ decreasing miR-155 expression was in a dose- and time-dependent manner. miR-155 could reduce the basal and AngⅡ-promoted ERK1/2, mTOR signaling pathway, while miR-155 inhibitor could elevate the above effect. Rap, U0126 and miR-155 could inhibit AngⅡ-attenuated expressions of SM22α, α-SM-actin and meanwhile inhibit AngⅡ-enforced expression of OPN. Conclusion: miR-155 could inhibit mice AngⅡ-promoted VSMC phenotype switching which might be via inhibiting the activations of mTOR and ERK1/2.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) and BMSCs-derived exosomes have similar functions, but the regulatory mechanism underlying the release of exosomes is still unclear. OBJECTIVE: To investigate the role of GW4869, an inhibition of neutral sphingomyelinase 2, in the release of exosomes in BMSCs and the influence of GW4869 on BMSCs proliferation. METHODS: Rat BMSCs were divided into three groups: normal control group, 24-hour GW4869 treatment group and withdrawal of GW4869 for 24 hours group (24-hour GW4869 treatment followed by 24-hour successive culture with drug withdrawal). Cultured cells were collected to extract exosomes by ultracentrifugation. Western blot was used to detect exosome-associated proteins CD63 and tumor susceptibility gene 101 (TSG101). The concentration and size distribution of exosomes were measured using nanoparticle tracking analysis. BCA was used to test the level of total proteins in exosomes. Live cell imaging system was used to observe the influence of GW4869 on BMSCs proliferation. RESULTS AND CONCLUSION: (1) Western blot results showed that exosomes expressed marker proteins such as CD63, TSG101. (2) Findings from the nanoparticle tracking analysis confirmed that the size of released exosomes was about 114 nm. (3) Significantly reduced release of exosomes was found in the two treatment groups compared with the normal control group (P < 0.01), but there was no significant difference between 24-hour GW4869 treatment group and withdrawal of GW4869 for 24 hours group (P > 0.05). (4) No significant difference in the proliferation of BMSCs was found among the three groups (P > 0.05). To conclude, 24-hour W4869 can inhibit the release of exosomes by BMSCs and this inhibitory effect is still sustained within 24 hours after drug withdrawal. However, GW4869 has no influence on the proliferation of BMSCs.

Objective To evaluate the safety and efficacy of cefaze-done injection ( CZD) compared with cefazolin injection ( CZL) in the treatment of acute bacterial respiratory infections.Methods Eligible subjects were divided randomly to receive 2.0 g cefazedone injection or cefazolin injection twice a day for 7 to 14 days.Efficacy and safety evaluation were done in accordance with the clinical trial protocol.Results Two hundred and sixty patients in 11 hospitals were en-rolled, 126 in CZD group( trial) and 134 in CZL group( control).There were no statistical differences in basic conditions between two groups( P >0.05 ).Cure rates of CZD group and CZL group were 95.5% and 94.9% in PPS ( P>0.05 ).Bacteria clearance rates of CZD group and CZL group were100% and 91.7% in BPPS and the total cure rates of CZD group and CZL group were 94.4% and 91.7% in BPPS, respectively ( P>0.05).Ten out off 126 patients in CZD group and 14 out off 134 in CZL group developed adverse events( AE ).Six and eleven events in CZD group and CZL group
were evaluated to be related with study drugs.One case in CZL group developed severe AE , which was considered not related with study drug.Conclusion Cefazedone injection is safe and effective in the treatment of respiratory infections.

OBJECTIVETo investigate the mechanisms of baicalin on anti-cerebral ischemic through observing the effect of baicalin on human brain microvascular endothelial cell under the glucose deprivation combined with hypoxia condition.

METHODSHuman brain microvascular endothelial cells (HBMVECs) cultured in vitro were divided into the following groups: normal group, model group, baicalin high dose group, baicalin middle dose group, baicalin low dose group, nimodipine group. The kits were used to detect the cell viability, leakage rate of lactate dehydrogenase (LDH), Na(+)-K(+)-ATPase, Ca(2+)-ATPase, the concentration of Ca2+ in each group, and apoptosis rates of each group were analyzed by flow cytometry (FCM).

RESULTSCompared with the normal group, the cell viability, Na(+)-K(+)-ATPase and Ca(2+)-ATPase in model group decreased significantly (P < 0.01, P < 0.05). But the leakage rate of LDH, Ca2+ in cells and apoptosis rates increased remarkably (P < 0.01). Compared with the model group, the cell viability, Na(+)-K(+)-ATPase and Ca(2+)-ATPase increased obviously (P < 0.01, P < 0.05) in baicalin high dose group. But the leakage rate of LDH and Ca2+ in cells in baicalin high dose group decreased significantly comparing with that of model group (P < 0.05, P < 0.01). And the reduction of intracellular Ca2+ was superior to that of nimodipine group. Meanwhile, the apoptosis rates decreased significantly in both baicalin high and middle dose groups (P < 0.01).

CONCLUSIONBaicalin could improve the cell viability of HBMVECs under the glucose deprivation combined with hypoxia condition. And the mechanisms were related with improving the energy metabolism, inhibiting intracellular calcium overload and decreasing the apoptosis rate of cells further.

Apoptosis ; Brain ; blood supply ; Calcium ; metabolism ; Capillaries ; cytology ; Cell Hypoxia ; Cell Survival ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Flavonoids ; pharmacology ; Glucose ; chemistry ; Humans