Objective:To determine the prevalence of Salmonella paratyphi C phage ( SPC-P1 ) in dif-ferent Salmonella serovars and to identify the integration sites in host genome. Methods: Based on the complete genome of SPC-P1 in S. paratyphi C RKS4594, 6 pairs of primers were designed and used to amplify the fragments of SPC-P1 in 11 S. typhi, 11 S. paratyphi A, 12 S. paratyphi B and 23 S. para-typhi C strains. At the same time, 100 complete genomes of Salmonella including 20 serovars available in National Center for Biotechnology Information ( NCBI) database were downloaded and aligned by Mauve 2. 3. 1 to determine the prevalence of SPC-P1 in these serovars. Primers were designed according to the integration sites of SPC-P1 in the genome of RKS4594 , and used to amplify ten strains having SPC-P1 in the genome. The PCR products were sequenced to investigate the integration sites of SPC-P1. Results:SPC-P1 was widely distributed in S. paratyphi C genome. In the study, 14 strains had all 6 fragments and 2 strains had 3-5 fragments. All the amplified fragments showed expected sizes. In contrast, in the ge-nomes of S. typhi, S. paratyphi A and S. paratyphi B, no or only 1-2 fragments could be amplified, and the sizes were smaller than expected. The results from Mauve showed that only in the genome of S. choleraesuis, which was a close relative of S. paratyphi C, there existed an almost complete genome of SPC-P1. The insertion site of SPC-P1 in all the ten S. paratyphi C strains tested was between pgtE and yfdC genes. Conclusion:SPC-P1 is a unique virulence factor of S. paratyphi C. It may play roles in the host range and pathogenicity of S. paratyphi C.

BACKGROUND: The principal deterrent to the success for hematopoietic stem cell transplantation (HSCT) is the complications after transplantation. The complications are associates with the conditioning regimens in the early stage. The highly-effective preparative regimens of proper dose and low-toxicity are the key to the successful HSCT.OBJECTIVE: To evaluate the curative effects and regimen related toxicity (RRT) of high-dose alkylating-agent-based chemotherapy as conditioning regimens for HSCT in the patients with hematological malignancies.DESIGN: Controlled study with observation.SETTING: Department of Hematology, Affiliated Hospital of Xuzhou Medical College.PARTICIPANTS: A total of 45 patients with leukemia and lymphoma hospitalized at Affiliated Hospital of Xuzhou Medical College from July 1997 to February 2006 were enrolled, including 31 males and 14 females. The median age was 31 years (from 7 to 52 years). The median course was 8 months (from 5 to 17 months) until transplantation.METHODS: Totally 45 patients with leukemia and lymphoma approached or got complete remission were treated by bone marrow transplantation and peripheral blood stem cell transplantation with preparative regimens of high-dose alkylating-agent-based chemotherapy. RRT was graded according to Bearman proposal, from grade 0 (no toxicity) to grade Ⅳ (fatal toxicity). The period of hematopoietic reconstitution, the rates of complete remission and relapse and disease-free survival were statistically observed in transplant recipients.MAIN OUTCOME MEASURES: Occurrence of RRT as conditioning regimens.RESULTS: ①Five patients did not show any toxicity. The greatest toxicity of grade Ⅲ was uncommon (13%, 6/45). Most of the cases with RRT were in grade Ⅰ - Ⅱ and severe oases in grade Ⅲ were rare. In grade Ⅰ - Ⅱ, stomatocace and gastrointestinal toxicity were common respectively of 73% (33/45) and 51% (23/45) which were recovered in short time after treatment; Heart toxicity was rare and only in grade Ⅰ, most of which were tachyoardia and changes of ST-T shape. The increase of transaminase was common in the clinical manifestations of liver RRT except two cases of HVOD.There were four oases of HC, in which one was delayed. RRT on kidney, lungs and CNS was uncommon. ②Totally 43 patients engrafted gained hematopoietic reconstitution, 2 patients died of implant failure (4%). Within the median follow-up period of 37 (8-102) months, 10 patients relapsed, 5 patients died of transplantation-related complications and 28 patients were alive in a disease-free situation (62.2%). The cause of death within 100 days after transplantation was ordinal as acute graft-versus-host disease (GVHD), cytomegalovirus (CMV) interstitial pneumonia, disseminated infections,multiple organ failure and early relapses.CONCLUSION: Alkylating-agent-based conditioning regimens may be well tolerated with low toxicities for HSCT in leukemia and lymphoma.

BACKGROUND: Tumor necrosis factor-α (TNF-α) is one of important cytokines to promote the maturation of dendritic cells. Blockage of TNF-α action by binding with soluble tumor necrosis factor receptor 1 (sTNFR1) may arrest dendritic cells in an immature state and induce stable, long-term tolerance. OBJECTIVE: To construct the lentiviral vectors carrying sTNFR1 gene and investigate sTNFR1 expression in immature dendritic cells. METHODS: Total RNA of human peripheral blood mononuclear cells was taken as a template. The sTNFR1 gene fragment was amplified by RT-PCR, subcloned to the lentiviral vectors pXZ208, and ligated to the enhanced green fluorescent protein (eGFP) reporter gene to establish lentiviral vector, called pXZ9-sTNFR1. DNA sequencing was performed for lentiviral vector identification. Lentivirus was prepared by transfection of 293 FT cells with pXZ9-sTNFR1. Viral titer was determined by eGFP expression. C57BL/6 mouse bone marrow-derived dendritic cells were in vitro cultured with low-dose granulocyte-macrophage colony stimulating factors and interleukin 4. On day 5 of culture, immature dendritic cells were transfected with pXZ9-sTNFR1 recombinant lentiviral supernatant, sTNFR1 transcription was detected by RT-PCR, sTNFR1 protein expression by Western blot analysis. Following sTNFR1 gene modification and lipopolysaccharide stimulation, the phenotype characteristics of dendritic cells were observed. RESULTS AND CONCLUSION: Recombinant plasmid pXZ9-sTNFR1 was successfully constructed. Twenty-four hours after 293 FT cell transfection, eGFP expression was observed and viral titer was over 10<'6> U/L. RT-PCR demonstrated that pXZ9-sTNFRl-transfected immature dendritic cells showed sTNFR1 positive expression. Western blot analysis revealed that sTNFR1 protein appeared in the immature dendritic cells and supernatant following 293 FT cell transfection. On day 5 of culture, dendritic cells expressed low level of class Ⅱ major histocompatibility complex (MHC Ⅱ), as well as CD40, CD86, CD80, molecules. However, following lipopolysaccharide stimulation, dendritic cells expressed high level of MHC Ⅱ, as well as CD40, CD80, and CD86, molecules, exhibiting the phenotype characteristics of mature dendritic cells. But after sTNFR modification, the expression level of MHC Ⅱ, as well as CD40, CD80, and CD86, molecules was not altered obviously. Lentiviral vectors carrying sTNFR1 gene and eGFP reporter gene were successfully constructed, and recombinant lentiviral plasmids with high titer were acquired. Following high efficacy of lentiviral gene transfection, immature dendritic cells stably express sTNFR1 mRNA and protein, which prevents immature dendritic cells from activation by exogenous lipopolysaccharide and maintains the immature state.

BACKGROUND: The primary qualification of peripheral blood stem cell transplantation (PBSCT) is the effective mobilization and harvesting of hematopoietic stem cells. The mobilization efficacy is closely related to the selection of high-efficacy low-toxicity regimen, the timing of mobilization and harvesting as well.OBJECTIVE: To investigate the efficacy of mitoxantone (MIT) combined with high-dose arabinosylcytosin (Ara-C),followed by granulocyte colony-stimulating factor (G-CSF) alone or combination of G-CSF and granulocyte-macrophage colony-stimulating factor (GM-CSF) on mobilizing PBSCs in patients with hematological malignancies and solid tumors.DESIGN: Controlled study with observation.SETTTNG: Department of Hematology, the Affiliated Hospital of Xuzhou Medical College.PARTICIPANTS: Forty-two patients with hematological malignancies and solid tumors admitted to Department of Hematology, Xuzhou Medical College from September 1998 to December 2006 were involved in this study. They were diagnosed according to FAB classification criteria and new WHO proposals. The involved patients, 25 male and 17 female, averaged 29 years ranging from 7 to 54 years and weighted (52±18) kg. Among them, 12 were patients with acute myeloblastic leukemia, 6 were patients with acute lymphoblastic leukemia (ALL), 1 was patient with chronic granulocytic leukemia (CGL) at chronic phase, 15 were patients with non-Hodgkin lymphoma (NHL), 4 were patients with Hodgkin lymphoma (HL), 2 was patient with multiple myeloma (MM), 2 were patients with advanced breast cancer. All the patients apprcached to or got complete remission after conventional chemotherapy. No tumor cell infiltration was observed in bone marrow cytological examination. The functions of the main organs such as heart, lung, liver and kidney,and so on, were normal. The patients underwent an average of 8-course chemotherapy before the mobilization. Informed consents of all the patients were obtained.METHODS: MIT was intravenously injected at 10 mg/(m2·d)for 2 to 3 days, then Ara-C was also intravenously injected at 2 g/m2 every 12 hours for 1 to 2 days. When white blood cell (WBC) count recovered from the lowest value, 5 to 7.5 μg/ (kg·d)G-CSF was applied in 20 patients for 3 to 5 days successively. And 5 to 7.5 μg/ (kg·d)G-CSF and 5 to 7 μ g/(kg·d)GM-CSF were applied in another 22 patients at 6:00 in the morning and in the evening, respectively. PBSCs harvesting started when WBC ＞ 2.5×109 L-1, especially when CD34+ cells≥ 1%,WBC was doubly increased. Autologous peripheral blood mononuclear cells (MNCs) were collected with CS3000 plus blood cell separator for detecting the level of CD34+ cells and T lymphocyte subsets. CFU-GM assays were performed in a methyl-cellulose-based clonogenic assay.① MNCs mixed with FITC-labeled CD34+, CD3 and CD8 monoclonal antibodies as well as CD4 PE-labeled CD monoclonal antibody at 4 ℃ for 30 minutes. 5×105 cells were determined, and CD3 and CD34+ levels, CD4/CD8 were determined by flow cytometer.Colony forming unit-granulocyte macrophage (CFU-GM) was determined with methyl cellulose. ② Related adverse reactions were observed after operation. ③ Aiming to different types of diseases,autologous PBSCs were back infused 36 to 48 hours after pre-disposal treatment. MNCs count and trypan-blue drying were done. Levels of CFU-GM and CD34+ cells were determined after unfreezing.MATN OUTCOME MEASURES: ① Changes in CD34+ cells and T lymphocyte subsets before and after mobilization. ② Postoperative related adverse reactions. ③ Back perfusion volume of autologous PBSCs (MNCs count, the number of CFU-GM and CD34+ cells).RESULTS: Forty-two involved patients participated in the final analysis. ① Changes in CD34+ cells and T lymphocyte subsets before and after mobilization: Without using hematopoietic growth factors (HGF), the percentage of CD34+ cells in peripheral blood of the patients was (0.054±0.032)%. After using G-CSF/GM-CSF treatment, it was (1.82±0.76)%,which was obviously increased compared with that of without using HGF (P ＜ 0.001). The CD34+ cells and CFU-GM yields of 22 patients in C-CSF plus GM-CSF combination group [(8.76±3.39)×106/kg, (3.52±1.33)×105/kg, respectively]were significantly higher than those of 20 patients in G-CSF alone group [(6.12±2.11)×106/kg, (2.03±1.07)×105/kg,respectively (P ＜ 0.05)]. There were no obvious changes of T lymphocyte subsets in the patients when using G-CSF/GM-CSF for some days except that CD34+ cells increased gradually (P ＞ 0.05). ② Postoperative related adverse reactions: Ⅱ to Ⅲ degree hair-loss was seen in all the patients. Blood platelets dropped to (54.43±26.14)×109 L-1 at different degrees. Infective fevers (37.8 ℃ to 41.0 ℃) occurred in 21 patients. But they were controlled in short term after antibiotics treatment. All the side effects of G-CSF and GM-CSF were mild and reversible, easily controlled with paracetamol or steroids. Bone pain (mainly in lumbosacral region) occurred in 13 patients when WBC went up quickly. ③ Back perfusion volume of autologous PBSCs: PBSCs were cryopreserved at -80 ℃ without program control for 2.0 to 6.5 months. The cell recovery rate was (88.7±7.4) %. Trypan blue exclusion rate was (92.1±5.5) %. The back perfusion volume of MNCs, CD34+ cells and CFU-GM yields were (5.21±2.44)×108/kg, (6.89±3.55)×106/kg, (2.58±2.33)×105/kg,respectively. ④Circulation blood volume were 10 to 16 L (end-point separation blood volume were all above trebling TBV). Hematopoiesis was well reconstituted in 40 patients received autologous PBSCT.CONCLUSTON: MIT and high-dose Ara-C chemotherapy combined with both G-CSF alone and G-CSF plus GM-CSF can safely and effectively mobilize autologous PBSCs, while G-CSF plus GM-CSF is superior to G-CSF alone.Large-volume leukapheresis is an important method to enhance the productive rate of stem cells and decrease the times of harvesting.

Objective To provide an experimental reference for clinic application of Jiazhu vascular ligating clips on ligation effects and the impact on the body in rabbits after laparoscopoic cholecystectomy.Methods 36 rabbits were randomly divided into 3 groups, normal control group, Jiazhu group and similar product control ( Hem-o-lok) group, 12 rabbits in each group.Except for normal control group, the rabbits were used gallbladder duct and gallbladder artery closed by vascular ligating clips and Hem-o-lok clips in Jiazhu group and similar product control group, respectively.The changes of blood biochemical, electrolytes, hematological parameters, coagulation, ligation effects and organ coefficients were observed at post-operation during 12 months.Results Compared with normal control group, the rabbit weight was reduced in the Jiazhu group at 3 days postoperatively ( P <0.01 ) , ALT level was increased at 1week postoperatively ( P <0.05), CREA and ALB levels were all significant decreased at 1~2 weeks postoperatively (P <0.05, P <0.01), NEUT numbers and TG content were significant increased at 2 weeks postoperatively ( P <0.05 ) , GLU content was significant decreased at 2 weeks and 1 month postoperatively ( P <0.01 ); LYM and RBC numbers were significant decreased at 1 month and 1 week postoperatively (P <0.05), respectively.PLT numbers were significant increased at 1~2 weeks postoperatively ( P <0.01) .While the rabbit weight was reduced in the Hem-o-lok group between 3 days and 1month postoperatively (P <0.05, P <0.01), and ALT level, NEUT and PLT numbers were increased at 1 week postoperatively (P <0.05, P <0.01), ALB and GLU contents were all significant decreased at 2 weeks postoperatively (P <0.01), TC content was significant increased at 1~2 weeks postoperatively (P <0.05).The rabbit weight, blood biochemical and hematological parameters in the Jiazhu group and Hem-o-lok group were all no significant differences with normal control group in the remaining time ( P >0.05 ) , and there were all no significant differences with electrolyte, coagulation and organ coefficients between each group (P >0.05).Conclusion Jiazhu vascular ligating clips are safe and reliable to close gallbladder duct and blood vessels, It shows that there are no differences in the validity and security between Jiazhu vascular ligating clips and Hem-o-lok clips.

Purpose: To construct an index system to evaluate the competencies of nurses in enhanced recovery after surgery (ERAS) programs and provide a scientific foundation for their training and assessment. Methods: Utilizing a literature review and semi-structured interviews, a preliminary indicator system was constructed. Based on the preliminary indicator system, a Delphi questionnaire was developed and utilized to achieve consensus among experts in two rounds of Delphi studies. The indicators were selected based on a mean importance score greater than 4 and a coefficient of variation less than .25. The weights of the indicators were calculated using the Analytic Hierarchy Process. Results: The study developed a system that evaluates the competencies of nurses involved in ERAS programs, offering a reference for their training and evaluation. The final index system includes 7 primary indicators, 20 secondary indicators, and 66 tertiary indicators. The primary indicators consist of competencies in the following components: 1) Direct clinical practice (20 items); 2) Expert coaching and guidance (9 items); 3) Consultation (6 items); 4) Research (7 items); 5) Leadership (11 items); 6) Collaboration (8 items); and 7) Ethical decision-making (5 items). Conclusion The developed competency evaluation index system is reliable and can serve as a foundation for the selection, training, and assessment of ERAS nurses.

Purpose: To construct an index system to evaluate the competencies of nurses in enhanced recovery after surgery (ERAS) programs and provide a scientific foundation for their training and assessment. Methods: Utilizing a literature review and semi-structured interviews, a preliminary indicator system was constructed. Based on the preliminary indicator system, a Delphi questionnaire was developed and utilized to achieve consensus among experts in two rounds of Delphi studies. The indicators were selected based on a mean importance score greater than 4 and a coefficient of variation less than .25. The weights of the indicators were calculated using the Analytic Hierarchy Process. Results: The study developed a system that evaluates the competencies of nurses involved in ERAS programs, offering a reference for their training and evaluation. The final index system includes 7 primary indicators, 20 secondary indicators, and 66 tertiary indicators. The primary indicators consist of competencies in the following components: 1) Direct clinical practice (20 items); 2) Expert coaching and guidance (9 items); 3) Consultation (6 items); 4) Research (7 items); 5) Leadership (11 items); 6) Collaboration (8 items); and 7) Ethical decision-making (5 items). Conclusion The developed competency evaluation index system is reliable and can serve as a foundation for the selection, training, and assessment of ERAS nurses.

Purpose: To construct an index system to evaluate the competencies of nurses in enhanced recovery after surgery (ERAS) programs and provide a scientific foundation for their training and assessment. Methods: Utilizing a literature review and semi-structured interviews, a preliminary indicator system was constructed. Based on the preliminary indicator system, a Delphi questionnaire was developed and utilized to achieve consensus among experts in two rounds of Delphi studies. The indicators were selected based on a mean importance score greater than 4 and a coefficient of variation less than .25. The weights of the indicators were calculated using the Analytic Hierarchy Process. Results: The study developed a system that evaluates the competencies of nurses involved in ERAS programs, offering a reference for their training and evaluation. The final index system includes 7 primary indicators, 20 secondary indicators, and 66 tertiary indicators. The primary indicators consist of competencies in the following components: 1) Direct clinical practice (20 items); 2) Expert coaching and guidance (9 items); 3) Consultation (6 items); 4) Research (7 items); 5) Leadership (11 items); 6) Collaboration (8 items); and 7) Ethical decision-making (5 items). Conclusion The developed competency evaluation index system is reliable and can serve as a foundation for the selection, training, and assessment of ERAS nurses.

Objective To explore the clinical application of transvaginal ultrasonography in the diagnosis of intrauterine adhesions .Methods 95 patients with suspected intrauterine adhesions were selected as the study subjects.All patients were treated with vaginal two -dimensional,three-dimensional ultrasound,and the results and hysteroscopy results were compared .The sensitivity,specificity,positive predictive value ,negative predictive value and accuracy of two -dimensional and three -dimensional ultrasonography were analyzed .Results Of 95 cases with suspected intrauterine adhesions ,vaginal two -dimensional ultrasound diagnosed 54 cases of intrauterine adhesions , including 30 cases of mild adhesion ,18 cases of moderate adhesion ,6 cases of severe adhesion ,22 cases of missed diagnosis or misdiagnosis.Compared with hysteroscopy ,the difference was statistically significant (χ2 =12.213,P=0.007).Three -dimensional ultrasound diagnosed intrauterine adhesions in 63 cases,including 20 cases of mild adhesion ,35 cases of moderate adhesion ,8 cases of severe adhesion ,only 7 cases of missed diagnosis or misdiagnosis . Compared with hysteroscopy ,the difference was not statistically significant (χ2 =0.630,P=0.889),suggested that the diagnosis of vaginal three -dimensional ultrasound and hysteroscopy results was consistent , and vginal three -dimensional ultrasound was superior than two -dimensional ultrasound , the difference was statistically significant (χ2 =8.848,P=0.003).The sensitivity of transvaginal two -dimensional ultrasonography in diagnosis of intrauterine adhesions was 67.65％,which of three-dimensional ultrasound was 89.71％,there was statistically significant differ-ence between the two groups (χ2 =9.861,P=0.002).The specificity of two-dimensional ultrasound was 70.37％, which of three-dimensional ultrasound was 92.59％,there was significant difference between the two groups (χ2 =4.418,P=0.036).The positive predictive value of two -dimensional ultrasound was 85.19％,which of three -dimensional ultrasound was 96.83％,there was statistically significant difference between the two groups (χ2 =5.040, P=0.025).The negative predictive value of two -dimensional ultrasound was 46.34％,which of three-dimensional ultrasound was 78.13％, there was statistically significant difference between the two groups (χ2 =7.583, P =0.006).The diagnostic accuracy of two -dimensional ultrasound was 68.42％,which of three -dimensional ultra-sound was 90.53％,there was statistically significant difference between the two groups (χ2 =14.228,P=0.000). Conclusion Transvaginal three-dimensional ultrasound in diagnosis of intrauterine adhesions is more accurate than two-dimensional ultrasound , and the result is consistent with hysteroscopy .Transvaginal three -dimensional ultra-sound can be used as the preferred method of intrauterine adhesions .

OBJECTIVE: To delineate cytogenetic and molecular abnormalities of a fetus carrying a de novo 46,X,der(X),t(X;Y)(p22.3;p11.2). METHODS: G-banded karyotyping and next-generation sequencing (NGS) were used to analyze the fetus, his father and sister. Single nucleotide polymorphism-based arrays (SNP-array), multiple PCR and fluorescence in situ hybridization (FISH) were utilized to verify the result. RESULTS: G-banded karyotyping at 320 bands showed that the fetus had a normal karyotype, while NGS has identified a 3.58 Mb microdeletion at Xp22.33 and a Y chromosomal segment of about 10 Mb at Yp11.32p11.2. With the sequencing results, high-resolution karyotyping at 550-750 bands level has determined the fetus to be 46,X,der(X)t(X;Y)(p22.3;p11.2). The result was confirmed by PCR amplification of the SRY gene, FISH and SNP-array assays. The karyotypes of his father and sister were both normal. His sister also showed no amplification of the SRY gene, and her NGS results were normal too, suggesting that the karyotype of the fetus was de novo. CONCLUSION Combined karyotyping, NGS, SNP-array, PCR and FISH assay can facilitate diagnosis of XX disorder of sex development.
Chromosomes, Human, X ; genetics ; Disorders of Sex Development ; genetics ; Female ; Fetus ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Translocation, Genetic