To investigate cytotoxic secondary metabolites of Micrococcus sp. R21, an actinomycete isolated from a deep-sea sediment (-6 310 m; 142 degrees 19. 9' E, 10 degrees 54. 6' N) of the Western Pacific Ocean, column chromatography was introduced over silica gel, ODS, and Sephadex LH-20. As a result, eight compounds were obtained. By mainly detailed analysis of the NMR data, their structures were elucidated as cyclo(4-hydroxy-L-Pro-L-leu) (1), cyclo(L-Pro-L-Gly) (2), cyclo( L-Pro-L-Ala) (3), cyclo( D-Pro-L-Leu) (4), N-β-acetyltryptamine (5), 2-hydroxybenzoic acid (6), and phenylacetic acid (7). Compound 1 exhibited weak cytotoxic activity against RAW264. 7 cells with IC50 value of 9.1 μmol x L(-1).
Animals ; Biological Factors ; chemistry ; isolation & purification ; metabolism ; pharmacology ; Cell Survival ; drug effects ; Macrophages ; cytology ; drug effects ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Mice ; Micrococcus ; chemistry ; genetics ; isolation & purification ; metabolism ; Molecular Structure ; Phylogeny ; RAW 264.7 Cells ; Seawater ; microbiology ; Secondary Metabolism

OBJECTIVETo explore optimal choice of surgical treatment and operative approach for closed complex tibial plateau fractures and its influencing factors.

METHODSFrom January 2003 to January 2011, 95 patients with closed complex tibial plateau fractures were estimated Schatzker V and Vl, and treated with three different surgical methods. The methods included single plate through anterolateral incision (Group A, 22 cases), double plates through inside and outside incisions (Group B, 36 cases), and double plates through antero-midline incisions (Group C, 37 cases). There were 56 males and 39 females, ranged the age from 19 to 57 years (averaged, 36.3 years), 50 cases in left, 45 cases in right. According to Schatzker classification, 51 cases were type V, 44 cases were VI. The data of operation time, intraoperative blood loss, complications (infectious of wound, necrosis, bad incision, collapse fracture, loosen of internal fixation, fracture failure)and recovery of function of lower limb joint were collected.

RESULTSThere were no significant difference among three groups in operation time (P > 0.05); blood loss in group A was obvious better than other groups (P < 0.05); collapse of joint surface and failure rate of internal fixation in group A was higher than other groups (P > 0.05); Merchant score after 1 year were higher in group B, C than group A. For lower limb function, 10 cases got excellent results, 5 good, 4 fair and 3 poor in group A; 21 cases got excellent results, 11 good, 3 fair and 1 poor in group B; 23 cases got excellent results, 11 good,2 fair and 1 poor in group C.

CONCLUSIONThe blood loss in group A was least, but fracture exposure and joint surface was not satisfactory, and stable fixation could not be achieved, the long-term result was not good. For fractures with double condyles and dislocated involved, double plates through inside and outside incisions or double plates through antero-midline incisions was suggested,which benefit good reduction of joint surface, stable fixation, and erlier exercise.

Adult ; Bone Plates ; Case-Control Studies ; Female ; Fracture Fixation ; adverse effects ; methods ; Fractures, Closed ; surgery ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery

OBJECTIVETo study the clinical characteristics and pathogens of invasive fungal infection in children.

METHODSThe clinical data of 104 children who suffered from invasive fungal infections between 2008 and 2012 was retrospectively reviewed.

RESULTSOf the 104 cases, 20 occurred in neonates, 48 in infants and 36 in preschool and school-aged children (old-aged children). Prematurity (70%), hyaline membrane disease (45%) and pneumonia (30%) were commonly comorbid in the neonate group. In addition, the percentage of cases receiving total parenteral nutrition was higher in the neonate group than in the other two age groups (P<0.01). Mechanical ventilation was more frequent in neonate and infant groups than in the old-aged children (P<0.01). Hematological malignancy was the most common underlying disease, and the percentage of children who had neutropenia and accepted chemotherapy was higher in the old-aged children than in the other two age groups (P<0.05). Lung infection was the most common (61.5%), followed by sepsis (14.4%) and intestinal tract infection (12.5%), while nervous system infections were found only in old-aged children. A total of 105 strains of fungi were isolated from the 104 patients, including Candida (n=90, 85.7%), Cryptococcus (n=6) and others (n=9). The most commonly isolated species was Candida albicans (n=52, 49.5%). Non-Candida albicans Candida accounted for 36.2% (n=38). The rate of susceptibility of Candida species to 5-fluorocytosine and amphotericin B was higher than fluconazole.

CONCLUSIONSInvasive fungal infections can occur in children at various ages. There are differences in the risk factors for invasive fungal infections between age groups. Candida species are the main pathogens of childhood invasive fungal infections, and both Candida albicans and non-Candida albicans Candida are common. Fluorocytosine and amphotericin B are sensitive antifungal agents for infections caused by Candida species.

Adolescent ; Child ; Child, Preschool ; Female ; Fungi ; drug effects ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Male ; Microbial Sensitivity Tests ; Mycoses ; drug therapy ; etiology ; microbiology ; Prognosis ; Risk Factors

OBJECTIVETo study common pathogens and their antibiotic susceptibility as well as clinical characteristics of neonatal pneumonia.

METHODSA retrospective study on neonatal pneumonia was performed. The study investigated antibiotic susceptibility of four common pathogens (339 strains) that caused neonatal pneumonia. Clinical characteristics of the newborns with pneumonia were analyzed. Of the 339 strains, 185 were isolated from bronchial secretions, 72 from blood samples, and 82 with positive results of both samples.

RESULTSFour hundred and seventy-four neonates with pneumonia presented positive results of bacterial culture. the most common pathogens Staphylococcus aureus (21.9%), Escherichia coli (19.2%), Klebsiella pneumoniae (19.0%) and Enterobacter cloacae (11.4%). The birth weight of newborns infected with Staphylococcus aureus was generally normal, and the time of hospital admission was later (after 24 hours of life). In contrast, the newborns with gram-negative bacterial infection, especially Klebsiella pneumoniae infection, had lower birth weights and early time of hospital admission (within 24 hours of life). Nearly more than 50% gram-negative bacteria were resistant to second, third and forth generation cephaloporins.

CONCLUSIONSGram-negative bacteria are predominant pathogens of neonatal pneumonia. Neonatal pneumonia caused by gram-negative bacteria is common in newborns with low birth weight and its onset time is relatively earlier. Gram-negative bacteria that cause neonatal pneumonia are highly resistant to cephaloporins.

Adult ; Birth Weight ; Drug Resistance, Bacterial ; Female ; Gram-Negative Bacteria ; drug effects ; isolation & purification ; Humans ; Infant, Newborn ; Male ; Maternal Age ; Microbial Sensitivity Tests ; Pneumonia ; microbiology ; Retrospective Studies

OBJECTIVETo establish a method for detection of reverse transcriptase region of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA), and to compare the pattern and frequency of drug-resistant mutations in the region between intrahepatic HBV cccDNA and serum HBV relax circle DNA (rcDNA).

METHODSHBV DNA were extracted from liver biopsy tissues of 20 patients with chronic hepatitis B. The RT region of HBV cccDNA was amplified by rolling circle amplification (RCA) followed by polymerase chain reaction (PCR) mediated by a pair of primers spanning across the gap region of HBV genome. The RT region of serum HBV rcDNA from the same patient was amplified by nested-PCR. The PCR products were directly sequenced and analyzed by Vector NTI Suite 8.0 and chromaslite 201 software. x2 test was used for statistical significance analysis of drug-resistant mutation occurrences between the HBV cccDNA and rcDNA.

RESULTSThe RT regions of HBV cccDNA were successfully amplified from liver tissues of all enrolled patients using the RCA plus PCR assay. Simultaneously, HBV the RT regions of rcDNA were amplified from these patients serum samples. Sequence analysis showed that the drug-resistant mutations were significantly more frequently detected in HBV rcDNA (40%) than in HBV cccDNA (10%) (P<0.05). Different mutational patterns were observed between the HBV cccDNA and rcDNA in a few cases.

CONCLUSIONThe RCA in combination with PCR is a practical method for the detection of drug-resistant mutation in the RT region of HBV cccDNA. Drug-resistant mutational patterns could be discrepant between HBV cccDNA and rcDNA.

DNA Primers ; genetics ; DNA, Circular ; genetics ; DNA, Viral ; genetics ; Drug Resistance, Viral ; genetics ; Genes, Viral ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Mutation ; Nucleic Acid Amplification Techniques ; methods ; Polymerase Chain Reaction ; methods ; RNA-Directed DNA Polymerase ; genetics ; Sequence Analysis, DNA

Objective To clone a gametocyte specific protein Pfgdv1 of Plasmodium falciparum,express and identify re?combinant Pfgdv1 protein in vitro. Methods PCR was performed to amplify Pfgdv1 from P. falciparum DNA which was got from the patient who was infected with P. falciparum,and the PCR product was inserted into pET28a(+)vector. pET28a?Pfg?dv1 recombinant plasmid was constructed and transformed into E. coli host BL21(DE3+). IPTG was used to induce the recombi?nant Pfgdv1 protein fused with His tag,and the protein was purified by His?NTA affinity chromatography. The recombinant pro?tein was identified by SDS?PAGE and Western blotting. Results The PCR product of Pfgdv1 gene was about 1.65 kb,meeting the expectation of predicted fragment size. The recombinant protein was about 67 kDa,which could be recognized by His?Tag monoclonal antibody. Conclusion The Pfgdv1 gene of P. falciparum is successfully cloned,and the recombinant Pfgdv1 pro?tein is expressed,thereby providing an opportunity for further study on transmission blocking vaccine.

objective To understand the baseline data of mortality from malignant tumor from 2006 to 2009 around Hongyanhe Nuclear Power Plant in Wafangdian City,Liaoning Province,so as to provide scientific basis for evaluating the impact of normal operation of nuclear power plant on the health of the residents nearby.Methods Thirty small towns near Hongyanhe Nuclear Power Plant were divided into 5 investigated areas according to the distances away therefrom(0.,10.,20-,30-,and 40-km).The data about from malignant tumor were obtained from the Center for Disease Control and Prevention of Wafangdian.The mortality distribution of difierent malignant tumors was analyzed,including the radiosensitive malignant tumors,especially leukemia,breast cancer,and thyroid gland cancer in different area,gender,and age groups.Results The mortality from malignant tumor was 151.97/105,and the standardized mortality rate(SMR)was 97.76/105.The mortality from malignant tumor among the males was 188.28/105(with the SMR of 116.76/105),and that among the females was 113.47/105(with the SMR of 75.89/105).with a sex ratio of 1.71.The first five cancers in the rank of death causes were lung,liver,stomach,colorectal,and esophageal cancers with mortality of 46.19/105,23.51/105,20.30/105,8.06/105 and 5.45/105,respectively.The mortality from mal.ignant tumor in the areas around the nuclear power plant from the near to the distant were 99.85/105, 137.40/105,138.73/105,156.30/105,and 154.16/105,respectively.The mortality from radiosensitive malignant tumors,leukemia,breast cancer,and thyroid gland cancer were 4.57/105,4.06/105,and 0.26/105,respectively.Conclusions Lung cancer and digestive tract malignant tumors are the main causes of death from malignant tumors in Wafangdian area before the nuclear power plant began to operate.There are no significant differences in the mortality distribution of malignant tumors among different areas,genders,and age groups.There are not significant differences in the mortality distribution of leukemia and breast cancer among different areas and age groups.

OBJECTIVETo investigate the genetic causes of azoospermia and severe oligozoospermia.

METHODSCytogenetic analysis and multiplex polymerase chain reaction(PCR) analysis were done on the 148 patients with azoospermia and serious oligozoospermia.

RESULTSEleven of the 148(7.4%) cases showed microdeletion of at least one STS. In fifteen STS of AZFa, AZFb,AZFd, AZFc, thirteen STS, eleven STS,two STS and one STS microdeletion were found in each case respectively, including two with 12 STS, five with 5 STS microdeletion.Seven cases had chromosomal morphologic changes(4.7%),four had deletion and one had deletion with translocation of long arm on Y chromosome. One had enlarged region one band two(q12) on long arm of Y chromosome and one had reciprocal translocation of autosomes.

CONCLUSIONThe findings indicated that AZF microdeletion and chromosomal abnormality should be important causes of male infertility.

Azoospermia ; genetics ; Chromosome Aberrations ; Chromosomes, Human, Y ; genetics ; Genetic Loci ; Humans ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Seminal Plasma Proteins ; genetics ; Sequence Deletion ; Sequence Tagged Sites

AIMTo investigate the spatial and temporal expression of germ cell nuclear factor (GCNF) in mouse and rat epididymis during postnatal period.

METHODSThe epididymal sections from different postnatal days were stained for GCNF by the indirect immunofluorescence technique and digital photographs were taken by a Carl Zeiss confocal microscope.

RESULTSGCNF was first detected on day 12 in mouse epididymis and day 14 in rat epididymis. The highest expression of GCNF was observed on day 35 in both mouse and rat epididymis. In adults, GCNF exhibited a region-specific expression pattern, i.e., it was expressed predominantly in the initial segment, caput and proximal corpus of rat epididymis and was abundant in the proximal corpus of mouse epididymis. GCNF could be found in the nuclei of the principal, apical, narrow, clear and halo cells.

CONCLUSIONGCNF may play an important role in epididymal differentiation and development and in sperm maturation.

Aging ; Animals ; DNA-Binding Proteins ; analysis ; Epididymis ; chemistry ; growth & development ; Fluorescent Antibody Technique, Indirect ; Male ; Mice ; Mice, Inbred BALB C ; Microscopy, Confocal ; Nuclear Receptor Subfamily 6, Group A, Member 1 ; Rats ; Rats, Sprague-Dawley ; Receptors, Cytoplasmic and Nuclear ; analysis ; Tissue Distribution