Adult ; China ; Humans ; Male ; Neisseria meningitidis, Serogroup C ; drug effects ; genetics ; isolation & purification

Bodily Secretions ; virology ; Humans ; Pharynx ; virology ; Pneumonia ; diagnosis ; virology ; Respiratory System ; virology

Objective To compare the genetic characteristics of mumps virus strain circulating in Beijing with vaccine strain and to preliminarily analysis the reasons of vaccine ineffectiveness. Methods The following methods were used: Isolation and identification of the mumps virus which had been circulating in Beijing, immunization history analysis, SH gene sequence analysis and comparison genotype homology with reference strains and analysis of the key amino acid sites of HN variation. Results In 38 mumps cases that virus had been isolated from, another seven cases were IgM negative. In 2007 and 2008, the positive rates on virus isolation, RT-PCR and IgMdecreased significantly, while the cases with immunization history had an increase. Cases without histories of vaccination had both higher positive rates on virus isolation and IgM. Thirty-eight strains belonged to F genotype virus, but vaccine strain was A genotype. The circulating viruses showed 5.6% sequence divergence on SH gene nucleotide and 16.0%-18.1% from vaccine strain. Conservative hydrophobic amino acids on SH protein of some Beijing strains had changed. For example, there were 6 strains, from No.8: L→F. The circulating viruses showed 2.3% sequence divergence on HN protein amino acid sequences and 4.2%-5.3% from vaccine strain. Amino acids sites, which deciding the ability of cross-neutralization of the Beijing strains and vaccine strains were different. At the 354 and 356 sites, all the Beijing strains were different from the vaccine strains. The N-glycosylation sites on HN of Beijing strains were also different from those on vaccine strains. Locations 464-466 appeared to be NCS on Beijing strain, but locations 464-466 were NCR on the vaccine strains. Another 18 unknown function amino acids sites of all Beijing strains were different from those on vaccine strains. Conclusion In recent years, genotype F became the main genotype of circulating strains in Beijing without genotype variation, but larger difference was found between them. There was a big difference between SH and HN protein of Beijing strains and vaccine strain, which might explain the ineffectiveness of the vaccine.

OBJECTIVETo identify wild measles virus and vaccine virus by detection nucleic acid of clinical samples from measles patients with immunization history circulating in Beijing through multiplex real-time fluorescent PCR technology.

METHODSFrom July 2011 to February 2012, 10 throat swabs and 15 urine specimens were collected from 16 suspected measles patients who were 8 - 9 months old infants with immunization history in Beijing. The specificity of multiplex real-time fluorescent PCR was firstly tested by measles vaccine virus, wild virus and other respiratory virus. Then the vaccine virus and wild virus were titrated and diluted to test the sensitivity of the PCR method. The result was then compared with it analyzed by PCR-RFLP method. Meanwhile, the clinical sample of the measles patients were tested and confirmed by sequencing method.

RESULTSThe primer-probe sets of Fam, Joe and Cy5 showed great specificity of measles virus, and could distinguish the measles vaccine virus and wild virus well. The sensitivity of this method to detect measles vaccine virus reached 0.1 CCID(50)/0.1 ml; and the sensitivity to wild virus reached 0.006 CCID(50)/0.1 ml; which were both higher than the sensitivity of PCR-RFLP method. Out of the 16 measles patients with vaccination history, 3 were negative and the other 13 all belonged to measles virus genotype A, and were confirmed to be vaccine virus by sequencing method.

CONCLUSIONMultiplex real-time PCR method is accurate, rapid and sensitive to identify measles vaccine virus and wild virus. The method could greatly help us to find measles patients and to identify the vaccine-related cases.

China ; epidemiology ; DNA Primers ; Humans ; Infant ; Measles Vaccine ; Measles virus ; classification ; genetics ; isolation & purification ; Multiplex Polymerase Chain Reaction ; RNA, Viral ; genetics ; Sensitivity and Specificity

OBJECTIVE: To induce hematopoietic progenitor/stem cells of umbilical cord blood to differentiate into mature megakaryocytes and platelets in vitro and to investigate the mechanism of production of platelets. METHODS: The CD34+ cells were sorted from umbilical cord blood by magnetic activated cell sorting (MACS) and then cultured in vitro with optimized medium to be differentiated into mature megakaryocytes and platelets. The cultured cells and the platelet-like particles were isolated from the culture and were checked by the fluorescence-activated cell sorter (FACS), immunohistochemistry assays, light microscope,electron microscope and platelet aggregation tests. RESULTS: The cultured megakaryocytes were detected with proplatelets and both the cultured cells and the platelet-sized particles were found to have the same structure with the normal megakaryocytes and platelets by light and electron microscope. The immunohistochemistry assays revealed the cultured cells expressed GP II b III a with a positivity of 95% which was a special antigen for platelets and megakaryocytes. Culture-derived platelet-sized particles aggregated in response to thrombin as the plasma derived-platelets did. The cultured platelets had the same positivity of CD41 as the platelets from platelet rich plasma. CONCLUSION The hematopoietic progenitor/stem cells can be induced to differentiate into purified and mature megakaryocytes and platelets. It provides a practical way to study the mechanism of platelets production.
Antigens, CD34 ; metabolism ; Blood Platelets ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Fetal Blood ; cytology ; metabolism ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Megakaryocytes ; cytology

Objective To study the changes of permeability of blood brain barrier (BBB) in rats after acute traumatic brain injury and explore its mechanism. Methods Ninety-six male Wistar rats were randomly divided into sham-operated group (S group) and traumatic brain injury group (TBI group).Each group was divided into 6 subgroups (3,6,12,24,48 and 72 h after trauma).Brain injury animal models were established according to Feeney' s method.Measurement of Evans Blue (EB) was performed at different time points after injury to measure the changes of BBB permeability.Brain water content was tested by wet-dry weighting method. Expression of tight junction protein Claudin-5 was detected by Western blotting. Results As compared with that in rats of the S group,the brain water content and content of EB in rats of the TBI group were significantly increased at each time point (P＜0.05); and in TBI subgroups,the brain water content and content of EB were begun to increase 3 h after TBI,reaching its peak level 24 h after TBI.As compared with that in rats of the S group,the expression of Claudin-5 in rats of the TBI group was significantly decreased at 12,24,48 and 72 h after TBI (P＜0.05); and in TBI subgroups,the expression of Claudin-5 was begun to decrease 6 h after TBI,reaching its lowest level 24 h after TBI. Negative correlations were noted between protein expression level of Claudin-5 and both the brain water content and content of EB (r=-0.994,P=0.000; r=-0.846,P=0.036); positive correlation was observed between the brain water content and content of EB (r=0.863,P=0.027). Conclusion The degree of brain injury and changes of permeability of BBB may be time dependent in rats after acute brain traumatic injury; and Claudin-5 may be correlated with the changes of BBB.

OBJECTIVETo explore a new method for repair of concurrent skin and nerve defect at palm and carpal on ulnar side.

METHODSFrom April 2000 to August 2009, five cases with concurrent skin and nerve defect at palm and carpal on ulnar side were reconstructed with free medial plantar flaps. Palmar nervous proprii defect at ulnar side of little finger was repaired by the first toe tibia nervous proprii in one case. The superficial branch of radial nerve was applied to repair the defect of ulnar nerve, as well as its deep or superficial branch in two cases. The superficial branch of radial nerve was also used to repair the defect of superficial branch of ulnar nerve, common palmar digital nerve of the fourth finger, Little finger ulnar palmar nervous proprii in one case. The dorsal branch of ulnar nerve was applied to repair the defect of superficial branch of ulnar nerve, common palmar digital nerve of the fourth finger, little finger ulnar palmar nervous proprii in one case. The vascular bundle of medial plantar flap was anastomosed with ulnar vascular bundle. The wounds at donor sites were covered with free skin grafts which were obtained from upper leg.

RESULTSAll the flaps and skin grafts were survived completely. The five patients were followed up for six months to four years with no muscular atrophy or claw hand deformity. The esthetic result was satisfied. The Sensory of flaps and fingers recovered to S3 to S3+. The two-point discrimination distance on flaps was range from 7 mm to 10 mm. The postoperative comprehensive evaluation was excellent in the cases whose superficial and deep branches of ulnar nerve were repaired.

CONCLUSIONSFree medial plantar flap is an effective method to repair concurrent skin and nerve defect at palm and carpal on the ulnar side.

Adult ; Female ; Foot ; surgery ; Free Tissue Flaps ; Hand Injuries ; surgery ; Humans ; Male ; Skin ; injuries ; Ulnar Nerve ; injuries ; surgery ; Wrist Injuries ; surgery ; Young Adult

Anti-Bacterial Agents ; pharmacology ; Bacterial Outer Membrane Proteins ; Bacterial Typing Techniques ; Electrophoresis, Gel, Pulsed-Field ; Microbial Sensitivity Tests ; Neisseria meningitidis, Serogroup B ; classification ; drug effects ; genetics ; Phylogeny

Purpose::Ischemia and hypoxia are the main factors limiting limb replantation and transplantation. Static cold storage (SCS), a common preservation method for tissues and organs, can only prolong limb ischemia time to 4 - 6 h. The normothermic machine perfusion (NMP) is a promising method for the preservation of tissues and organs, which can extend the preservation time in vitro by providing continuous oxygen and nutrients. This study aimed to evaluate the difference in the efficacy of the 2 limb preservation methods. Methods::The 6 forelimbs from beagle dogs were divided into 2 groups. In the SCS group ( n = 3), the limbs were preserved in a sterile refrigerator at 4 °C for 24 h, and in the NMP group ( n = 3), the perfusate prepared with autologous blood was used for the oxygenated machine perfusion at physiological temperature for 24 h, and the solution was changed every 6 h. The effects of limb storage were evaluated by weight gain, perfusate biochemical analysis, enzyme-linked immunosorbent assay, and histological analysis. All statistical analyses and graphs were performed using GraphPad Prism 9.0 one-way or two-way analysis of variance. The p value of less than 0.05 was considered to indicate statistical significance. Results::In the NMP group, the weight gained percentage was 11.72% ± 4.06%; the hypoxia-inducible factor-1α contents showed no significant changes; the shape of muscle fibers was normal; the gap between muscle fibers slightly increased, showing the intercellular distance of (30.19 ± 2.83) μm; and the vascular α-smooth muscle actin (α-SMA) contents were lower than those in the normal blood vessels. The creatine kinase level in the perfusate of the NMP group increased from the beginning of perfusion, decreased after each perfusate change, and remained stable at the end of perfusion showing a peak level of 4097.6 U/L. The lactate dehydrogenase level of the NMP group increased near the end of perfusion and reached the peak level of 374.4 U/L. In the SCS group, the percentage of weight gain was 0.18% ± 0.10%, and the contents of hypoxia-inducible factor-1α increased gradually and reached the maximum level of (164.85 ± 20.75) pg/mL at the end of the experiment. The muscle fibers lost their normal shape and the gap between muscle fibers increased, showing an intercellular distance of (41.66 ± 5.38) μm. The contents of vascular α-SMA were much lower in the SCS group as compared to normal blood vessels.Conclusions::NMP caused lesser muscle damage and contained more vascular α-SMA as compared to SCS. This study demonstrated that NMP of the amputated limb with perfusate solution based on autologous blood could maintain the physiological activities of the limb for at least 24 h.

OBJECTIVESTo detect the serum proteomic patterns by using SELDI-TOF-MS (surface enhanced laser desorption/ ionization-time of flight-mass spectrometry) technology and CM10 ProteinChip in colorectal cancer (CRC) patients, and to evaluate the significance of the proteomic patterns in the tumour staging of colorectal cancer.

METHODSSELDI-TOF-MS and CM10 ProteinChip were used to detect the serum proteomic patterns of 76 patients with colorectal cancer, among them, 10 Stage I, 19 Stage II, 16 Stage III and 31 Stage IV samples. Different stage models were developed and validated by support vector machines, discriminant analysis and time-sequence analysis.

RESULTSThe Model I formed by 6 protein peaks (m/z: 2759.58, 2964.66, 2048.01, 4795.90, 4139.77 and 37761.60) could be used to distinguish local CRC patients (Stage I and Stage II) from regional CRC patients (Stage III) with an accuracy of 86.67% (39/45). The Model II formed by 3 protein peaks (m/z: 6885.30, 2058.32 and 8567.75) could be used to distinguish locoregional CRC patients (Stage I, Stage II and Stage III) from systematic CRC patients (Stage IV) with an accuracy of 75.00% (57/76). The Model III could distinguish Stage I from Stage II with an accuracy of 86.21% (25/29). The Model IV could distinguish Stage I from Stage III with accuracy of 84.62% (22/26). The Model V could distinguish Stage II from Stage III with accuracy of 85.71% (30/35). The Model VI could distinguish Stage II from Stage IV with accuracy of 80.00% (40/50). The Model VII could distinguish Stage III from Stage IV with accuracy of 78.72% (37/47). Different stage groups could be distinguished by the two-dimensional scattered spots figure obviously.

CONCLUSIONThis method showed great success in preoperatively determining the colorectal cancer stage of patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Colorectal Neoplasms ; blood ; diagnosis ; pathology ; surgery ; Female ; Gene Expression Profiling ; methods ; Humans ; Male ; Middle Aged ; Neoplasm Proteins ; blood ; Neoplasm Staging ; Preoperative Care ; methods ; Protein Array Analysis ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods