How to get functional gene from uncultured-microbiology is the hotspot content of microbial ecology. What the most important is how to obtain the pure and integrated genomic DNA. An efficient, nonselective extraction method to gain chromosomal DNA from eight kinds of bacteria was introduced. Amount DNA released by hot-detergent gave the highest DNA yields from different G + and G- bacteria. Running 20 hours by PFGE mode, the size of total DNA is over 23kb. The pure DNA could be digested by Hind Ⅲ and used in PCR. The total environmental DNA also can be extracted from soil by the same method. As a result it showed a new way for the environmental DNA extraction.

Methods for studying the population diversity of microorganism in activated sludge usually require enrichment of bacterial genome.The efficient information on microbial species composition provided and shifted in diversity revealed are dependent on the effective DNA recovery technique.The method was based on washing by alkaline phosphate buffer and digestion with extended heating of the activated sludge suspension in the presence of lysozyme and freeze-thawing in high-salt-SDS buffer.The extraction was tested for four activated sludge differing in places and dates.The DNA fragment from all sludge was integrity.DNA yields ranged from 105 to 823 ?g/g sludge and were of sufficient purity for PCR-based 16S ribosomal DNA analysis and restriction digested.In general,all methods produced DNA pure were not enough for PCR amplification and libraries construction.As basis of experimental goals,the study provides an appropriate extraction method of microbial DNA in sludge.

OBJECTIVETo study the effects of L.F04, the active fraction of Lycopus lucidus, on erythrocytes rheological property so as to investigate its mechanism in promoting blood circulation and removing blood stasis.

METHODThe effects of L.F04 (used for treatment for 10 days in different dosages) on deformability, aggregation and membrane liquidity of erythrocytes (MLE) as well as whole blood apparent viscosity (eta(b)) were examined on the basis of rat model of blood-stasis syndrome induced by venous injection of high molecular weight dextran.

RESULTAs compared with the normal control group, the model group's RBC deformability and MLE were lower, and the aggregation of erythrocytes and eta(b) were higher. Compared with the model group, both L.F04 0.612 g/kg and 0.306 g/kg showed significant effect in improving deformability and inhibiting aggregation of red blood cells (RBC) and reducing blood viscosity. The trend of improving MLE was also shown.

CONCLUSIONL.F04 could significantly improve the abnormal rheological property of erythrocytes.

Animals ; Blood Viscosity ; drug effects ; Dextrans ; pharmacology ; Drugs, Chinese Herbal ; administration & dosage ; Erythrocyte Aggregation ; drug effects ; Erythrocyte Deformability ; drug effects ; Hemorheology ; Hemostasis ; drug effects ; Lycopus ; Male ; Rats ; Rats, Wistar ; Space Flight

The purpose of this study was to determine the changes of pathogens in hematological ward and susceptibility of patients received chemotherapy to antibiotics. The pathogens were taken from blood, urine and sputum of patients who accepted chemotherapy from years 2001 to 2005, then were isolated and identified. The susceptibility test was performed by disk diffusion method. The results showed that the total of 418 strains were detected. Gram-negative bacteria were the most common of nosocomial infection. Pseudomonas aeruginosa, Enterobacter cloacae, E. coli account for the most of Gram negative- bacteria infection and most resistant to broad-spectrum penicillin, Acinetobacter baumannii showed a trend of increase. The ratios of gram positive bacteria and fungi were increased slowly, mainly as Enterococcus and Candida. Enterococcus is the most common cause of Gram-positive bacterial infection. Vancomycin resistance did not occur. It is concluded that Gram-negative bacteria are main cause of nosocomial infection in patients with hematological malignancies. Gram positive bacteria and fungi had been more frequent. Strains resistant to antimicrobial agents increase.
Cross Infection ; epidemiology ; microbiology ; Drug Resistance, Bacterial ; Gram-Negative Bacteria ; drug effects ; isolation & purification ; Gram-Negative Bacterial Infections ; epidemiology ; microbiology ; Hematologic Diseases ; microbiology ; Hematologic Neoplasms ; microbiology ; Humans ; Microbial Sensitivity Tests

Objective To evaluate the MRI features of central neurocytoma (CNC). Methods Thirteen patients with CNC (7 males and 6 females, aged 18-28), admitted to our hospital fiom January 2007 to January 2010, were chosen; preoperative plain and contrast enhanced MRI scans were performed on these 13 patients and 1 also received CT scan. The clinical and imaging data and pathological features of these patients were retrospectively analyzed. Results The largest tumor enjoyed its maximum diameter of 3.2-8.5 cm; the CNC in 12 patients located at the lateral ventricle and that in 1 patient at the parenchyma of the left frontal and temporal lobes. Lobulated solid tumors were noted in these lesions,showing T1 isointense and T2 hyperintense in MRI; some multiple, scattered cystic lesions showed stronger T1 and T2 signal than the former ones; vascular "flow void" signal was shown in 6 patients in T2WI, and slightly weak T1 signal was shown in 8. All the tumors demonstrated inhomogeneous enhancement and the tumor vessels in 6 patients were noted under contrast MRI. Focal calcification was found. Conclusion CNC in the lateral ventricle enjoys typical location; considering the age of patients, non-contrast and contrast MRI findings, correct diagnosis of CNC can be made; however, CNC outside the lateral ventricle is hard to diagnose.

Objective： The current study was designed to investigate the effect of heat shock protein 70(HSP70) on stability of mutant p53 protein. Methods： Retroviral recombinant expressing antisense HSP70 RNA was constructed and transfected into MCF7/Adr breast cancer cells. The existence of foreign DNA was identified by PCR method and the HSP70 protein level was determined by Western blot analysis. The half-life of mutant p53 protein(mtp53) was measured by p53 stability assay. Results： The stable expressing strain(MAp70) from transfected cells was obtained through G418 selection. The foreign DNA in transfectant cells were confirmed by PCR, and the repression rate of HSP70 protein was 42％ . The half-life of mutant p53 in MAp70 cell was 12 hours, significantly lower than that of the control cells. Conclusion： Antisense HSP70 RNA can decrease the HSP70 protein level and significantly increase instability of mutant p53 protein in MCF7/Adr breast cancer cell.

Analysis of serological and genetic characteristics on 2009 swine-origin influenza A (H1N1) virus (S-OIV) isolated from four patients with severe disease in Shenzhen were performed. Microneutralization assay showed that the neutralizing antibody titers of the infected patients did not exceed 1 : 20 in a short term post infection, which could not neutralize the viruses efficiently. Hemagglutination inhibition (HI) tests confirmed that the antigenicity of S-OIV from the patients was distinct from the seasonal influenza A virus, but similar to the reference strains of S-OIV. Phylogenetic and molecular analysis showed that S-OIV from the patients still belonged to the classical swine lineages and did not have the genetic characteristics of highly pathogenic influenza virus. Several amino acid residue mutations on HA protein were detected, which seemed not to affect the virulence and pathogenicity of the viruses. Further, A His 275 Tyr mutation on NA protein of a virus strain was detected, which induced the oseltamivir resistance of the virus.
Amino Acid Sequence ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Child ; Child, Preschool ; China ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; enzymology ; genetics ; isolation & purification ; Influenza, Human ; immunology ; virology ; Male ; Molecular Sequence Data ; Mutation ; Phylogeny ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; Young Adult

OBJECTIVETo investigate the severity related influencing factor and treatment strategy of severe acute pancreatitis with early organ dysfunction.

METHODSFrom July 2007 to December 2008, 167 patients with severe acute pancreatitis were treated in the Surgical Department of Ruijin Hospital. The relationships between the happening of early organ dysfunction and outcome of the patients were observed, with operative or nonoperative treatment strategy.

RESULTSAmong 167 patients, 68 patients have early organ dysfunction, in which 39 with single organ dysfunction and 29 with multiple organ dysfunction. The early organ dysfunction were involved in 47.1% in cardiovascular system, 35.3% in lung and 29.4% in kidney. Aging (P < 0.05) and higher APACHE II score (P < 0.05) predicted a poor prognosis, which were benefit from early operation.

CONCLUSIONSThe mortality of the patients with SAP is related to age, and the degree of organ dysfunction as well. In the first phase of the disease, the selection of operation depends on the trends and the degree of early organ dysfunction before infected necrosis happens, with the aid of SOFA score as a scale.

Acute Disease ; Aging ; Humans ; Multiple Organ Failure ; Pancreatitis ; diagnosis ; Prognosis

Objective To explore the effect of pedicled omental flap with Da Vinci surgical system for breast reconstruction,so as to facilitate the application of robot-assisted surgery in the breast reconstruction after breast cancer(BC) surgery.Methods A luminal BC patient,administrated in March 2017,received Da Vinci robot-assisted filling with pedicled omental flap after extensive lumpectomy and sentinel lymph node biopsy.Blood supply,wound healing,cosmetic outcome,complications and satisfaction of the patient were evaluated.Results The wound healed well,with no obvious scar.The reconstructed breast had desirable volume and looked symmetrical against the other side.The cosmetic outcome was excellent and the patient was satisfied with the surgery.No recurrence or metastasis was found during the twomonth post-operational follow-up.Conclusion Da Vinci robot-assisted breast reconstruction with pedicled omental flap has the advantages of clear vision,precise movement and flexible machinery joint.Breast conservation and autologous reconstruction with this technique substantially improves the psychological and social well-being of the patient.

BACKGROUNDAn implantable cardioverter-defibrillator (ICD) has been suggested for heart failure patients for primary prevention of sudden cardiac death. However, few data have been reported on the application of ICD as primary prevention of sudden cardiac death in China. We evaluated the value of primary prevention ICD therapy in Chinese patients with heart failure.

METHODSThirty-four patients at an average age of (60.2 +/- 13.7) years seen in Peking Union Medical College Hospital were treated with ICD implantation for primary prevention of sudden cardiac death from November 2005 to July 2009. Single-chamber ICDs were implanted in 16 (47.0%) cases, and dual-chamber or cardiac resynchronization therapy defibrillators in 18 (53.0%) cases. The patients had an average left ventricular ejection fraction of (26.9 +/- 5.5)% (11% to 35%), of which 18 (53.0%) patients had ischemic cardiomyopathy and 16 (47.0%) patients had non-ischemic cardiomyopathy. All patients were followed up at three months after the implantation and every six months thereafter or when prompted by an ICD event.

RESULTSThere were five (14.7%) deaths, including two of heart failure and three with a non-cardiac course, during an average follow-up of (15.0 +/- 11.9) months. Forty-one ICD therapy events were recorded, including 19 (46.3%) appropriate ICD therapies in six patients and 22 (53.7%) inappropriate ICD therapies in four patients with single chamber leads. Inappropriate ICD therapies were mainly due to supraventricular tachyarrhythmias, especially atrial fibrillation. Patients with ischemic cardiomyopathy and non-ischemic cardiomyopathy did not differ in the incidence of either appropriate or inappropriate therapy.

CONCLUSIONSICD for primary prevention of sudden cardiac death in China prevents patients from arrhythmia death. Relatively high incidence of inappropriate therapies highlights the importance of an atrial lead.

Aged ; Death, Sudden, Cardiac ; prevention & control ; Defibrillators, Implantable ; adverse effects ; Female ; Heart Failure ; mortality ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome