Klebsiella pneumoniae(K.pneumoniae)is an important pathogen that can cause severe hospital-and community-acquired infections.To systematically investigate its methylation features,we determined the whole-genome sequences of 14 K.pneumoniae strains covering varying serotypes,multilocus sequence types,clonal groups,viscosity/virulence,and drug resistance.Their methy-lomes were further characterized using Pacific Biosciences single-molecule real-time and bisulfite technologies.We identified 15 methylation motifs[13 N6-methyladenine(6mA)and two 5-methylcytosine(5mC)motifs],among which eight were novel.Their corresponding DNA methyl-transferases were also validated.Additionally,we analyzed the genomic distribution of GATC and CCWGG methylation motifs shared by all strains,and identified differential distribution pat-terns of some hemi-/un-methylated GATC motifs,which tend to be located within intergenic regions(IGRs).Specifically,we characterized the in vivo methylation kinetics at single-base resolu-tion on a genome-wide scale by simulating the dynamic processes of replication-mediated passive demethylation and MTase-catalyzed re-methylation.The slow methylation of the GATC motifs in the replication origin(oriC)regions and IGRs implicates the epigenetic regulation of replication initiation and transcription.Our findings illustrate the first comprehensive dynamic methylome map of K.pneumoniae at single-base resolution,and provide a useful reference to better understand epigenetic regulation in this and other bacterial species.

Objective    To evaluate the changes of left ventricular structure and function by echocardiography and its grading of left ventricular diastolic function in patients with mitral valve prolapse treated by minimally invasive mitral valve repair. Methods    By retrospective analysis, 37 patients including 25 males and 12 females aged 53.49±11.02 years with mitral valve prolapse who underwent minimally invasive mitral valve repair were as an operation group, and 34 healthy persons including 19 males and 15 females aged 54.26±8.33 years matched by age and sex were selected as a control group. Ultrasound parameters of every participant were routinely collected before operation, 1 month, 3 months, 6 months and 1 year after operation, and left ventricular diastolic function was graded. The ultrasound parameters between the two groups were compared. Results    The diameters of left ventricular end systolic and diastolic phase, left atrial diameter and left ventricular volume in the operation group were significantly smaller than those before operation. The diameters of left ventricle and left atrium after operation were significantly shorter than those before operation, but they were still larger than those of the control group. The ejection fraction value decreased significantly at one month after the operation and then returned to normal level. The incidence of left ventricular diastolic dysfunction at 6 months and 1 year after operation was significantly lower than that before operation (P<0.05). Conclusion    Minimally invasive repair for patients with mitral valve prolapse can significantly improve systolic and diastolic functions of left ventricle while reconstructing left atrial and left ventricular structures.

Objective To explore the feasibility and technical tips of the reconstruction of defects around the knee using the reverse-flow thigh flaps based on different branches of the lateral circumflex femoral artery ( LCFA) .A classification method of these flaps was proposed .Methods According to the characteristics of the defects and the LCFA arborizing condition , an optimal reverse-flow thigh flap was chosen for defect reconstruction .Results From 2010 to 2016, 23 patients underwent defect reconstruction using the reverse-flow thigh flaps .The flaps were classified into four different types according to the origin of the perforating vessels on which the flap was based , including the descending ( typeⅠ) , oblique ( typeⅡ) , rectus femoris ( type Ⅲ) , and transverse branches ( type Ⅳ) .In our series of patients , 7 type Ⅰflaps, 10 typeⅡ, 5 typeⅢ, and 1 typeⅣwere included.The reverse flow from the rectus femoris branch was combined and a dual pedicle was formed in 5 patients.The flap size ranged from 8 cm ×7cm to 24 cm × 12 cm.The perforating vessels included septocutaneous in 7 cases and musculocutaneous in the remaining 16.The pedicle length and distance from the pivot point to the superolateral border of the patella were 8.5 cm to 25 cm (mean, 17 cm) and 5 cm to 16 cm (mean, 11.48 cm) respectively.Pre-transfer donor site expansion was conducted in 5 patients.The donor sites were managed by direct closure in 18 cases, free skin grafting in 4, and transferring a pedicled deep inferior epigastric artery perforator flap in the remaining 1.All flaps survived after the surgery except one in which the venous congestion of the flap occurred postoperatively , resolved after using conservative treatment .The patients were followed for 1 to 48 months, tumor recurrence was not noticed and all patients were satisfied with the reconstructive outcomes .Conclusions LCFA-based classification system of the reverse-flow thigh flaps would provide surgeons more options when a defect around knee needs to be reconstructed .

Objective To explore the feasibility and technical tips of the reconstruction of defects around the knee using the reverse-flow thigh flaps based on different branches of the lateral circumflex femoral artery ( LCFA) .A classification method of these flaps was proposed .Methods According to the characteristics of the defects and the LCFA arborizing condition , an optimal reverse-flow thigh flap was chosen for defect reconstruction .Results From 2010 to 2016, 23 patients underwent defect reconstruction using the reverse-flow thigh flaps .The flaps were classified into four different types according to the origin of the perforating vessels on which the flap was based , including the descending ( typeⅠ) , oblique ( typeⅡ) , rectus femoris ( type Ⅲ) , and transverse branches ( type Ⅳ) .In our series of patients , 7 type Ⅰflaps, 10 typeⅡ, 5 typeⅢ, and 1 typeⅣwere included.The reverse flow from the rectus femoris branch was combined and a dual pedicle was formed in 5 patients.The flap size ranged from 8 cm ×7cm to 24 cm × 12 cm.The perforating vessels included septocutaneous in 7 cases and musculocutaneous in the remaining 16.The pedicle length and distance from the pivot point to the superolateral border of the patella were 8.5 cm to 25 cm (mean, 17 cm) and 5 cm to 16 cm (mean, 11.48 cm) respectively.Pre-transfer donor site expansion was conducted in 5 patients.The donor sites were managed by direct closure in 18 cases, free skin grafting in 4, and transferring a pedicled deep inferior epigastric artery perforator flap in the remaining 1.All flaps survived after the surgery except one in which the venous congestion of the flap occurred postoperatively , resolved after using conservative treatment .The patients were followed for 1 to 48 months, tumor recurrence was not noticed and all patients were satisfied with the reconstructive outcomes .Conclusions LCFA-based classification system of the reverse-flow thigh flaps would provide surgeons more options when a defect around knee needs to be reconstructed .

Objective To compare eradication rates,safety and compliance of minocycline quadruple and tailored therapies in patients retreated for Helicobacter pylori (H.pylori) infection.Methods Between January 2014 and June 2014,135 patients with dyspepsia (18-70 years) and H.pylori infection after at least one previous eradication treatment at a tertiary hospital were randomly assigned to a 10-day treatment with minocycline quadruple therapy versus tailored triple regimen of PPI,amoxicillin and a third antibiotic.In the group of tailored therapy,medications were adjusted based on clarithromycin sensitivity and cytochrome P450 isoenzyme 2C19 genotype.Eradication status was assessed 4-12 weeks after treatment.Results Although H.pylori eradication rates were higher in the minocycline therapy group than that in the tailored therapy group in intention-to-treat [84.1％ (95％ CI 75.0％-93.2％) vs 75.8％ (95％ CI 65.1％-86.5％),P =0.245] and per-protocol [88.3％ (95％ CI 80.3％-96.3％) vs 79.7％ (95％CI 69.7％-89.7％),P =0.197] analyses,the differences between the two groups were not statistically significant.The incidence of adverse effects and compliance between the two groups were also comparable.Conclusions The tailored therapy in this study had a poor eradication efficacy in the retreated patients with H.pylori infection.Minocycline quadruple therapy achieved a relatively satisfactory eradication efficacy and may be an alternative choice for the retreatment of H.pylori infection.Clinical trial registration Chinese Clinical Trial Registry,ChiCTR-TRC-13003975.

Objective To explore the clinical significance of typical reflux symptoms in the diagnosis of gastroesophageal reflux disease (GERD).Methods Consecutive patients older than 16 years,who initially visited department of gastroenterology at clinic of Peking University Third Hospital from May 9,2012 to Dec 31,2012,were required to complete a self-reported GERD questionnaire.Upper endoscopy was performed in some selected patients.Results A total of 18 987 patients were enrolled with a response rate of 91.5％.The prevalence of symptom-defined GERD was 13.6％ (2 579/18 987).A total of 4 357 (22.9％) patients underwent the upper endoscopy,and the diagnostic rates of reflux esophagitis,Barrett's esophagus,peptic ulcer disease,and upper gastrointestinal malignancy were 13.1％ (572/4 357),1.8％ (78/4 357),10.5％ (456/4 357),and 1.7％ (75/4 357),respectively.The incidence of reflux esophagitis was 22.7％ (216/951) in patients with reflux symptoms and 10.5％ (356/3 406) (P ＜0.001) in patients without reflux symptoms,2.7％ (26/951) and 1.5 ％ (52/3 406),respectively (P =0.013) for Barrett's esophagus; 6.8％ (65/951) and 11.5％ (391/3 406),respectively (P＜0.001) for peptic ulcer disease; 1.7％ (16/951) and 1.7％ (59/3 406),respectively (P =0.917) for upper gastrointestinal malignancy.Conclusions GERD is one of the major diseases at gastroenterology clinic.Typical reflux symptoms suggest a diagnosis of GERD.But some patients with peptic ulcer disease or upper gastrointestinal malignancy can also present typical reflux symptoms.Upper endoscopy is valuable to avoid the misdiagnosis of other disorders.

BACKGROUNDHelicobacter pylori (Hp) is a common and potentially curable cause of gastric mucosa lesion. This study investigated the relationship of Hp infection with histological changes in gastric mucosa and gastric cancer in Hp-positive patients compared with Hp-eradication patients followed up for ten years.

METHODSFrom an initial group of 1 006 adults, 552 Hp-positive subjects were randomly assigned to a treatment group (T; n = 276) or a placebo group (P; n = 276). In the randomized, double-blind, placebo-controlled, parallel trial, T group subjects received oral doses of omeprazole, amoxicillin and clarithromycin for 1 week; those in the P group received a placebo. One month after treatment ended, a 13C urea breath test was performed, and Hp was undetectable in 88.89% of the T group. All subjects were followed at 1, 5, 8, and 10 years after treatment, with endoscopy and biopsies for histological examination.

RESULTSGastric mucosa inflammation was significantly milder in the T group than that in the P group one year after Hp eradication and this persisted for 10 years. Glandular atrophy and intestinal metaplasia (IM) had deteriorated in both groups during ten years. However, the increased score of glandular atrophy at both the gastric antrum and corpus, and IM only at the gastric antrum, in the P group was more obvious than that in the T group. During the 10 years, 9 patients were diagnosed with gastric cancer (2 in the T group; 7 in the P group; P = 0.176). When mucosal atrophy was absent at the gastric antrum and corpus when entering the study, the incidence of gastric cancer in the P group (n = 6) was much higher than that in the T group (n = 0, P = 0.013).

CONCLUSIONSHp eradication may significantly diminish and help halt progression of gastric mucosal inflammation and delay the development of IM and atrophy gastritis. Hp eradication is helpful for reducing the risk for gastric cancer, especially in the early stage of Hp infection.

Adult ; Aged ; Amoxicillin ; therapeutic use ; Anti-Bacterial Agents ; therapeutic use ; Clarithromycin ; therapeutic use ; Double-Blind Method ; Female ; Follow-Up Studies ; Gastric Mucosa ; drug effects ; pathology ; Gastritis, Atrophic ; diagnosis ; drug therapy ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; pathogenicity ; Humans ; Male ; Middle Aged ; Omeprazole ; therapeutic use ; Stomach Neoplasms ; diagnosis ; prevention & control

OBJECTIVEThis study aimed to evaluate the effects of the microarc oxidation surface on cell morphology and cytoskeleton.

METHODSPure titanium with a diameter of 15 mm and a thickness of 1 mm was divided into four groups: grooved surfaces (group G); sandblasted surfaces (group SB); grooved microarc oxidation surfaces (group GMAO); and sandblasted microarc oxidation surfaces (group SBMAO). Osteoblast cells were cultured in each group. The morphology and proliferation of the cells on the titanium surface were observed by scanning electron microscope (SEM). The cytoskeleton was evaluated by laser scanning confocal microscope (LSCM).

RESULTSThe osteoblasts were inoculated after 12 h; these osteoblasts then spread along the surface of the titanium plate. In GMAO and SBMAO groups, the osteoblasts converged in the hole. The actin fibers in each group were clearly visible. In particular, the actin fibers in GMAO and SBMAO groups were arranged in parallel and formed bundles that extended into the holes.

CONCLUSIONAfter microarc oxidation, the titanium surface significantly affected the morphology and cytoskeleton of osteoblasts.

Actins ; Cytoskeleton ; Humans ; Microscopy, Electron, Scanning ; Microtubules ; Osteoblasts ; Oxidation-Reduction ; Surface Properties ; Titanium

OBJECTIVETo study the efficacy and safety of Naoxintong capsule treatment of stroke recovery with Qi-deficiency and blood-stasis syndrome (cerebral infarction), and to compared the non-inferiority analysis with the positive drug Tongxinluo capsule.

METHODTaking Tongxinluo capsules as control, randomized, double-blind, controlled, multi-center clinical experiments were studied. The evaluating indexes included the decrease of integral value of stroke patients, changes in traditional Chinese medicine, the improvement of the patient viability status (disability level), Chinese stroke scale (CSS), activities of daily living (DAL) scale and barthel index (BI ) points.

RESULTThe total effect of the two groups, Chinese and other symptoms, showed no significant statistical significance.

CONCLUSIONNaoxintong capsule stroke recovery, with Qi-deficiency and blood-stasis syndrome (cerebral infarction) has a therapeutic effect, and more secure.

Activities of Daily Living ; Adult ; Aged ; Capsules ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Qi ; Stroke ; drug therapy ; Treatment Outcome