OBJECTIVETo explore the hematopoietic stem cell distribution and lymphocyte proliferation and differentiation in recipient mice after allogeneic bone marrow transplantation (allo-BMT).

METHODSBALB/c (H-2(d)) mice were total body irradiated 5.5 Gy x 2 by (137)Cs and then transplanted with bone marrow cells from GFP transgenic C57BL/6J (H-2(b)) mice. The femur, spleen, Peyer patches, thymus, liver and peripheral blood of the host were collected on days 3, 7, 21, 35 and 70 post transplantation, and their sections were observed by fluorescent microscopy. The green fluorescent cells were counted with FACS. The phycoerythrin (PE) labeled antibodies to CD4, CD8 and B220 were used for sorting T and B lymphocytes.

RESULTS(1) On day 3 and day 7 after allo-BMT, there were (1.06 +/- 0.02)% and (76.60 +/- 1.80)% of donor's green bone marrow cells in host's spleen respectively, whereas only (0.37 +/- 0.06)% and (39.70 +/- 5.38)% in the bone marrow, respectively. (2) In bone marrow and other organs of 21 day-old chimerism mice, over 60% cells were of donor origin. (3) There were (0.36 +/- 0.04)% donor's bone marrow cells lodging at host's Peyer patches, similar to that in bone marrow.

CONCLUSION(1) The engrafted allogeneic hematopoietic stem cell can move into spleen, bone marrow, Peyer patches and thymus. The spleen is the main lodging place of the engrafted cells early after all-BMT. (2) The majority of cells in chimerism mice immunologic organs were of donor origin. (3) Peyer patches is another lodging place early after allo-BMT.

Animals ; Bone Marrow Cells ; immunology ; Bone Marrow Transplantation ; immunology ; Lymphocytes ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Tissue Donors

OBJECTIVETo analyze the genetic polymorphism, distribution of haplotypes, common and well-documented (CWD) and rare alleles of high-resolution HLA-A, B and DRB1 alleles by analysis from hematopoietic stem cell (HSC) donors in Jiangsu Han Chinese.

METHODSPCR-sequence-based typing and PCR-sequence specific oligonucleotide probes methods were applied for HLA-A, B and DRB1 high-resolution genotyping of 3238 unrelated healthy donors of hematopoietic stem cells in Jiangsu branch of Chinese National Marrow Donor Program registry.

RESULTS46 alleles of HLA-A,85 HLA-B and 51 HLA-DRB1 locus were found. The frequencies of the most common alleles were A * 11:01 (16.52%), B * 13:02 (11.60%) and DRB1 *07:01 (15.78%). That of the most common haplotype was A * 30: 01-B * 13: 02-DRB1 * 07: 01 (8.87%). 40 alleles of HLA-A,77 alleles of HLA-B, and 47 HLA-DRB1 alleles of HLA-DRB1 were CWD, which account for 99. 8% of total number of samples, and a few rare alleles not reported in Chinese population were found.

CONCLUSIONThe results of high-resolution, CWD and rare alleles showed the characteristics of HLA distribution in Jiangsu Han population, which may be useful for finding HLA matched unrelated donors, as well as for HLA correlation with population genetics and disease association studies.

Alleles ; Asian Continental Ancestry Group ; genetics ; Genotype ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DRB1 Chains ; genetics ; Haplotypes ; Hematopoietic Stem Cells ; Humans ; Tissue Donors

OBJECTIVETo compare the safety and efficacy between two different surgical approaches for thoracoscopic esophagectomy including left lateral decubitus position and prone position.

METHODSFrom January 2008 to December 2009, 88 patients who underwent thoracoscopic esophagectomy were enrolled in this study. Among them, 52 patients were placed in decubitus position and 36 patients were placed in prone position.

RESULTSNo conversion to thoracotomy occurred in either group. The operative time was shorter in the prone group than that in the decubitus group (70 ± 20 min vs. 82 ± 17 min, P<0.01). Blood loss during operation was less in the prone group(100 ± 52 ml vs. 139 ± 54 ml, P<0.01). More lymph nodes were harvested from chest in the prone group(12.2 ± 6.2 vs. 8.6 ± 4.3, P<0.01). There was no significant difference between the two groups in morbidity.

CONCLUSIONThoracoscopic esophagectomy in prone position is associated with better exposure of surgical filed, shorter operative time, less blood loss, and more extensive lymph node dissection as compared to decubitus position.

Aged ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Female ; Humans ; Male ; Middle Aged ; Posture ; Prone Position ; Retrospective Studies ; Thoracoscopy ; Treatment Outcome

OBJECTIVETo explore the safety of video-assisted thoracoscopic esophagectomy for esophageal carcinoma.

METHODSFrom January 2005 to March 2012, 260 patients with esophageal carcinoma received thoracoscopic esophagectomy (TE group), while 322 patients underwent conventional open esophagectomy (OE group). Operative procedures, perioperative complications, reoperation, readmission to intensive care unit (ICU), and perioperative mortality were compared between the two groups.

RESULTSCompared with OE group, TE group possessed less thoracic operative time [(105±30) min vs. (112±41) min, P=0.000], less blood loss [(95±48) ml vs. (107±44) ml, P=0.002], shorter postoperative hospital stay [(14.3±7.5) d vs. (16.9±9.5) d, P=0.000] and more lymph node harvest from thorax [(13.5±5.0) vs. (11.6±4.7), P=0.000]. The total perioperative complication rate was lower in TE group than that of OE group (34.6% vs. 45.0%, P=0.011), as well as perioperative mortality (0.8% vs. 3.4%, P=0.032). Lower rate of readmission to ICU (5.4% vs. 10.6%, P=0.024) was found in the TE group as compared to the OE group, while the reoperation rate was comparable (1.5% vs. 2.5%, P=0.425).

CONCLUSIONThoracoscopic esophagectomy is advantageous than open procedure in terms of surgical safety.

Aged ; Esophageal Neoplasms ; surgery ; Esophagectomy ; adverse effects ; methods ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Thoracoscopy ; adverse effects ; methods ; Video-Assisted Surgery

Objective:To analyze effects of esophageal suspension on left recurrent laryngeal nerve lymph node dissection ,operation duration ,and perioperative morbidity in minimally invasive esophageal surgery .Methods:Clinical data of 145 patients with esophageal cancer treated by minimally invasive surgery in Zhongshan Hospital ,Fudan University ,from Jan . to Dec .2015 were retrospectively analyzed ,including 71 cases of esophageal suspension in left recurrent laryngeal nerve lymphadenectomy and other 74 cases of conventional methods .Results:Compared with the conventional method group ,left laryngeal recurrent nerve lymph node dissection number of the esophageal suspension group (2 .55 ± 0 .20 vs 1 .46 ± 0 .22 , P< 0 .05) increased significantly ,operation time ([262 .60 ± 6 .44] min vs [265 .60 ± 6 .17] min) and dissection time ([9 .90 ± 0 .34] min vs [9 .60 ± 0 .36] min) did not increase significantly ,and the related complications such as irreversible injury in recurrent laryngeal nerve and thoracic duct injury did not increase .Conclusions:Esophageal suspension in minimally invasive esophageal surgery helps improve the quality of left recurrent laryngeal nerve lymphadenectomy without increasing operation duration and perioperative morbidity .

Objective:To analyze the activity of lithium chloride (LiCl) against influenza virus A (H3N2) in human non-small cell lung cancer cells (A549).Methods:Different concentrations of LiCl were incubated with A549 cells, and the cytopathic effect (CPE) was observed after 24 hours, and the effect of LiCl on cell activity was determined by CCK-8 method. After H3N2 (MOI=1) infected A549 cells, different concentrations of LiCl were added and incubated for 24 hours, and the viral load was measured by real time/reverse transcription quantitative polymerase chain reaction (RT-qPCR), and the CPE was observed, and the viral titer was determined. Different concentrations of LiCl were incubated with A549 at 37 ℃ and 5% CO 2 for 2 hours, virus was added and incubated for 24 hours, and the viral load was determined by RT-qPCR. LiCl, H3N2 and A549 were incubated at 4 ℃ for 1 hour, 35 ℃, 5% CO 2 for 24 hours, and viral load was determined by RT-qPCR. H3N2 and A549 were incubated at 4 ℃for 1 hour, then different concentrations of LiCl were added, incubated at 35 ℃ with 5% CO 2 for 24 hours, and the viral load was determined by RT-qPCR. After H3N2 infected A549 cells, different concentrations of LiCl were added and incubated for 24 hours, and the viral RNA load and viral titer of the supernatant and cells were measured, respectively, and then the corresponding ratios of the supernatant and the cells were calculated. After H3N2 (MOI=10) and BV (MOI=1) infected A549 cells, different concentrations of LiCl were added for 24 h, and the viral load was determined by RT-qPCR. Results:When the concentration of LiCl was<50 mmol/L, the cell viability of A549>90%. Different concentrations of LiCl could significantly reduce the viral load of H3N2 ( P<0.000 1), and the CPE of the LiCl treatment group was more dose-dependent than that of the control group. LiCl did not inhibit viral replication by affecting the cell itself; Different concentrations of LiCl significantly inhibited the entry of H3N2 into A549 ( P<0.000 1), and also had a certain inhibitory effect on the adsorption of A549 cells ( P<0.1). LiCl did not affect the assembly and release of H3N2 ( P>0.05), and it was also found that LiCl had a broad spectrum of antiviral effects against multiple influenza virus strains ( P<0.000 1). Conclusions:LiCl may exert antiviral effect by inhibiting the adsorption and entry of H3N2 into A549 cells and the replication of H3N2 in A549 cells, which provides a data reference for the prevention and treatment of viral infection by LiCl.

OBJECTIVETo explore the clinical feature and genetic diagnosis for Smith-Magenis syndrome (SMS).

METHODThe clinical data, including craniofacial anomalies, physical and mental status were analyzed. Routine and high resolution G-banding was performed to analyze the karyotype of the patient and her parents, and array comparative genomic hybridization (array CGH) was used to detect small chromosome anomaly.

RESULTA-two-year old girl was sent to our clinic for mental retardation and cardiac malformation. Some sleep problems were reported by parents, including difficulties falling asleep, shortened sleep cycles. She also had some neurobehavioral symptoms including hyperactivity and self-injurious behaviors head-banging. She had distinctive craniofacial features including low hairline, frontal bossing, a broad face, broad nasal bridge, a tented upper lip, prognathism, low-set ears and high-vaulted arch. She had moderate mental retardation. Cardiac findings included ventricular septal defect, atrial septal defect, overriding aorta and pulmonary hypertension. Primary ventriculomegaly was seen in magnetic resonance imaging (MRI). Routine karyotype analysis showed a karyotype of 46, XX. However, high resolution karyotype analysis showed a suspected partial deletion of the short arm of chromosome 17. Array comparative genomic hybridization (array CGH) finely mapped the deletion to a 3.8 Mb region on 17p11.2. The molecular karyotype was then ascertained as 46, XX.arr17p11.2(16543655-20374751)×1dn. The parents had normal karyotypes.

CONCLUSIONSmith-Magenis syndrome is a multisystem disorder characterized by developmental delay and mental retardation, distinctive craniofacial features, sleep disturbance and behavioral problems. Array comparative genomic hybridization (array CGH) finely mapped the deletion on 17p11.2.

Child, Preschool ; Chromosome Deletion ; Chromosomes, Human, Pair 17 ; Female ; Humans ; Intellectual Disability ; Karyotyping ; Smith-Magenis Syndrome ; diagnosis ; genetics

BACKGROUNDNonsyndromic hearing loss (NSHL) is highly heterogeneous, in which more than 90 causative genes have currently been identified. DFNA5 is one of the deafness genes that known to cause autosomal dominant NSHL. Until date, only five DFNA5 mutations have been described in eight families worldwide. In this study, we reported the identification of a novel pathogenic mutation causing DFNA5 deafness in a five-generation Chinese family.

METHODSAfter detailed clinical evaluations of this family, the genomic DNA of three affected individuals was selected for targeted exome sequencing of 101 known deafness genes, as well as mitochondrial DNA and microRNA regions. Co-segregation analysis between the hearing loss and the candidate variant was confirmed in available family members by direct polymerase chain reaction (PCR)-Sanger sequencing. Real-time PCR (RT-PCR) was performed to investigate the potential effect of the pathogenic mutation on messenger RNA splicing.

RESULTSClinical evaluations revealed a similar deafness phenotype in this family to that of previously reported DFNA5 families with autosomal dominant, late-onset hearing loss. Molecular analysis identified a novel splice site mutation in DFNA5 intron 8 (IVS8+1 delG). The mutation segregated with the hearing loss of the family and was absent in 120 unrelated control DNA samples of Chinese origin. RT-PCR showed skipping of exon 8 in the mutant transcript.

CONCLUSIONSWe identified a novel DFNA5 mutation IVS8+1 delG in a Chinese family which led to skipping of exon 8. This is the sixth DFNA5 mutation relates to hearing loss and the second one in DFNA5 intron 8. Our findings provide further support to the hypothesis that the DFNA5-associated hearing loss represents a mechanism of gain-of-function.

Adult ; Deafness ; genetics ; Exons ; genetics ; Female ; Hearing Loss ; genetics ; Hearing Loss, Sensorineural ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; genetics ; Young Adult

Objective To investigate the effect of percutaneous cardiac intervention (PCI) on the clinical prognosis in aged patients with coronary heart disease (CHD) involving multiple coronary arteries who obtained complete revascularization (CR) or incomplete revascularization (ICR) after PCI.Methods A total of consecutive 257 aged patients (＞75 years old) with confirmed CHD that involved multiple coronary arteries,who were admitted to authors' hospital during the period from January 2015 to September 2015 to receive PCI,were enrolled in this study.Based on the complete revascularization (CR) or incomplete revascularization (ICR) after PCI,the patients were divided into CR group and ICR group.The basic clinical data,PCI parameters and the occurrence of major adverse cardiovascular and cerebrovascular events (MACCE) in hospitalization days and in the follow-up period were compared between the two groups.Results CR group included 171 patients (66.53％) and ICR group included 86 patients (33.47％).The hypertension history,diabetes history,diagnosis of acute non-ST segment elevation myocardial infarction or myocardial occlusion disease at admission,chest tightness,palpitation and other discomfort symptoms occurring in one and 3 months after PCI,and re-hospitalization rate in ICR group were significantly higher than those in CR group (P＜0.05).No statistically significant differences in the incidence of MACCE during hospitalization days and at one,3 and 6 months after PCI existed between the two groups (P＞0.05).Conclusion In aged patients with CHD that affects multiple coronary arteries,ICR does not increase the risk of MACCE after PCI,although the re-hospitalization rate and the incidence of postoperative discomfort symptoms will be increased.The long-term prognosis needs to be further studied.

Objective To develop a rapid,sensitive and specific real time reverse transcription PCR for detecting and identifying human metapneumovirus. Methods The Hmpv-L gene of human metapneumovirus was chosen as target gene,the primers and TaqMan probe were designed,and the PCR reaction was optimized systematically. The total RNA was extracted from respiratory specimens,and reverse transcription was performed through random primer. The cDNA was detected by using real time PCR. The specificity,sensitivity and reproducibility of real time PCR were estimated. The real time PCR was applied to detect 180 clinical respiratory specimens. Results The human metapneumovirus can be detected using real time reverse transcription PCR accurately and quickly,and the sensitivity was 1 copy/μl.The coefficient of variation of intra-assay and inter-assay wasless than 5%. Among those 180 specimens,28(15.56%) were positive for human metapneumovirus,the clinical diagnoses for these 28 patients were pneumonia ( 15.60%,17/109) and bronehiolitis ( 15.49%,11/71 ) .21 positive specimens were from patients under 2 years of age,and 6 positive specimens were from patients between 2 and 5 years of age,only 1 positive specimens was from patients over 5 years. Conclusion It is demonstrated that real time reverse transcription PCR is a reliable,accurate and feasible assay for human metapneumovirus,whieh has become one of the most important pathogens induced acute respiratory infections in pediatric patients.