Expression and Clinical Significance of Peripheral Blood T cell JAK2/STAT3 mRNA in Chronic Idiopathic Thrombocytopenic Purpura / 中国实验血液学杂志

OBJECTIVE@#To detect the expression of JAK2/STAT3 mRNA in peripheral blood T cells from the patients with chronic idiopathic thrombocytopenic purpura（CITP）, and to explore the relationship between JAK2/STAT3 mRNA and CITP.@*METHODS@#CITP group and healthy control group were set in this study, The JAK2/STAT3 mRNA expression level in peropheral blood T cells of 2 groups was detected with the RT-PCR and agarose gel electrophoresis.@*RESULTS@#JAK2 mRNA expression level in CITP group was significantly higher than that in control group（P<0.01）, the STAT3 mRNA expression level in CITP group was also higher than control group（P<0.01）, The JAK2/STAT3 mRNA expression level of CITP patiants increased obviously compared with control group.@*CONCLUSION@#The expression level of JAK2/STAT3 mRNA increases signficanlty in chronic ITP patients, which involves in pathogenesis of CITP.

Combination of R848 and Poly（I:C）promotes antigen presentation by DC and enhances killing effect of CTL on lung adenocarcinoma A549 cells / 中国病理生理杂志

AIM: To investigate the effect of R848(a Toll-like receptor 7/8 agonist)combined with poly-inosinic:polycytidylic acid [Poly(I:C),a Toll-like receptor 3 agonist] on dendritic cell(DC)maturation,and the killing effect of DC-induced cytotoxic T-lymphocytes(CTL)on human lung adenocarcinoma A549 cells.METHODS:Mononu-clear cells were isolated from human peripheral blood and induced to differentiate into DC.The whole-cell lysate of A549 cells,namely tumor cell lysate(TCL), was used as antigen.R848 combined with Poly(I:C)was used as adjuvant to stimulate the DC.DC surface markers were analyzed by flow cytometry.The DC stimulated by antigen was co-cultured with T-lymphocytes for 7 d to induce CTL.The culture supernatant and CTL were collected.The levels of interleukin-12(IL-12)p70,interferon-γ(IFN-γ)and tumor necrosis factor-α(TNF-α)in the supernatant were measured by ELISA.The CTL and A549 cells were co-cultured for 16 h,and the cytotoxicity was observed by LDH assay.RESULTS:The expres-sion of CD83 and CD80 on the DC surface,and the secretion of IL-12 p70 in DC-R848+Poly(I:C)group were significant-ly increased compared with DC-TCL group(P<0.01).In addition,the cytotoxicity of CTL for A549 cells in DC-R848+Poly(I:C)group was significantly enhanced compared with DC-TCL group(P<0.01).The secretion levels of IFN-γand TNF-αin DC-R848+Poly(I:C)group were significantly elevated compared with DC-TCL group(P<0.01).CONCLU-SION:R848 combined with Poly(I:C)significantly promotes DC maturation and activation, and enhances the antigen-presenting effect of DC and the cytotoxicity of DC-induced CTL.

Immunogenicity of insulin producing cells differentiating from human umbilical cord mesenchymal stem cells / 中国组织工程研究

BACKGROUND: Human umbilical cord mesenchymal stem cells (hUC-MSCs) have low immunogenicity and it is unclear whether insulin producing cells (IPCs) that differentiate from hUC-MSCs have low immunogenicity. OBJECTIVE:To investigate the immunogenicity of IPCs differentiating from hUC-MSCs in vitro and after IPCs transplantation into the host. METHODS: (1) The hUC-MSCs were induced to differentiate into IPCs according to the modified scheme. Flow cytometry assay was used to detect the immunophenotype and apoptotic rate of IPCs in a cytotoxicity test. (2) Cell counting kit-8 was used to detect the proliferative capacity of human peripheral blood mononuclear cells in the one-way mixed lymphocyte assay. (3) The IPCs were then transplanted into the abdominal cavity and left renal capsule of mice, and then the infiltration of immune cells was detected by flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: The IPCs highly expressed HLA-ABC and lowly expressed HLA-DR, CD40 and CD80. The apoptosis rate of IPCs increased with the increase of pre-sensitized splenocytes in the cytotoxicity test. In the one-way mixed lymphocyte assay, IPCs inhibited the proliferation of human peripheral blood mononuclear cells when the target ratio was 10:1 and 50:1. After IPCs transplantation, the number of lymphocytes was increased in the transplanted site. In summary, our results show that IPCs that differentiate from hUC-MSCs maintain low immunogenicity in vitro,but have some immunogenicity after transplantation into the host due to microenvironment changes.

Efficacy on analgesia with electric stimulation of long-term retaining needle after laparoscopic cholecystectomy / 中国针灸

<p><b>OBJECTIVE</b>To observe the clinical effect of electric stimulation of long-term retaining needle on analgesia after laparoscopic cholecystectomy (LC) and the impacts on the post-surgical flatus time.</p><p><b>METHODS</b>Under static absorptive composite general anesthesia, 90 cases of LC were randomized into three groups, 30 cases in each one. In the control group, the analgesia was not applied after LC. In the analgesia-pumper group, the patient controlled intravenous analgesia (PCIA) was used. In the needle-retaining group, the electric acupuncture stimulator was used. The needles were inserted transversely at Riyue (GB 24), Qichong (ST 30) and Yanglingquan (GB 34) and fixed with sterile sticker. Separately, in 8 h and 24 h after surgery, the electric acupuncture stimulation with disperse-dense wave, 2 Hz/100 Hz frequency was applied continuously for 30 min. Visual analogue scale (VAS), adverse reactions such as vomiting and nausea and the postoperative flatus time in 2, 4, 8, 12, 24 and 36 h after surgery were observed and recorded in the three groups.</p><p><b>RESULTS</b>In 2, 4, 8, 12 and 24 h after surgery, VAS scores in the needle-retaining group and the analgesia-pumper group were all lower than those in the control group (P < 0.05, P < 0.01). The analgesia effect at the above time points in the needle-retaining group was better than that in the analgesia-pumper group (all P < 0.05). There was not adverse reaction in the needle-retaining group. But there were 3 cases of somnolence, 6 cases of nausea and 3 cases of vomiting in the analgesia-pumper group, and 2 cases of nausea and 1 case of vomiting in the control group. The flatus time was quite earlier in the needle-retaining group as compared with the other two groups [(14.77 +/- 4.99) h vs (18.50 +/- 4.22) h, P < 0.01; (14.77 +/- 4.99) h vs (18.17 +/- 4.69) h, P < 0.05].</p><p><b>CONCLUSION</b>The electric stimulation of long-term retaining needle is safe and effective in analgesia after LC. It avoids the adverse reactions of analgesics and promotes postoperative flatus.</p>

Effect of p21Ras in vitro intervention on the phenotype of airway smooth muscle cells from asthmatic rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>An important physiological feature of asthma is the phenotypic change of airway smooth muscle cells (ASMCs), but the precise mechanisms behind the ASMCs' change remains unknown. Our study assessed whether p21Ras can directly modulate the phenotype of ASMCs.</p><p><b>METHODS</b>Rat ASMCs were treated with FTP III, a highly specific p21Ras inhibitor. ASMCs were identified via immunocytochemistry. The ultrastructure of cells was observed by electron microscopy, and the expression of α-actin was evaluated by Western blotting analysis. The levels of IL-6 and RANTES were measured by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>It was observed that ASMCs in asthma exhibited a proliferative/secretory phenotype and were larger, denser and had many pseudopods, as well as increased signs of secretory organelles. Additionally, the level of α-actin, a marker of ASMCs, was reduced in asthmatic ASMCs and the secretion of IL-6 and RANTES was increased. When FTP III was added to asthmatic ASMCs it induced a contractile phenotype, with increased α-actin levels and reduced secretion of IL-6 and RANTES.</p><p><b>CONCLUSIONS</b>It appears that p21Ras induces asthmatic ASMCs to a proliferative/secretory phenotype, but its inhibitor FTP III, can significantly reverse this phenotype. The role of p21Ras in the ASMCs may be a new target for asthma treatment.</p>

Qualitative research on the family management and family needs of children with asthma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>A qualitative research was conducted to investigate the problems on the family management of asthmatic children and the needs for family health services in order to provide basis for family-centered care.</p><p><b>METHODS</b>Fifteen caregivers of children with asthma were interviewed with open-ended questions. The collected data were studied using Colaizzi's seven-step method of phenomenological analysis.</p><p><b>RESULTS</b>The problems of family management and the needs for family health services were shown as follows: insufficient knowledge to prevention and treatment of asthma, poor compliance, ignoring psychological effects of asthma on children, a family's failure to cope with the distress and financial burden.</p><p><b>CONCLUSIONS</b>It is important to provide asthma education and prevention program for caregivers and encourage them to participate in the design of medical program for asthmatic children. Individual asthma education and guides are also necessary for caregivers.</p>

Clinical significance of enrichment and detection of circulating tumor cells in NSCLC patients with immunomagnetic beads / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the feasibility and clinical significance of detection of circulating tumor cells (CTCs) in peripheral blood of NSCLC patients by lung cancer cell immunomagnetic enrichment and detection kit.</p><p><b>METHODS</b>Four groups of patients (Group A: 18 cases with stage I lung cancer; Group B: 33 cases with stage II - IV lung cancer; Group C: 20 cases with benign pulmonary diseases; Group D: 20 healthy volunteers) were enrolled for detection of CTCs using the lung cancer cell enrichment and detection kit developed by ourselves, and compared with the results obtained using simple ICC method and the detection of CK19 mRNA and LUNX mRNA using nested PCR as control.</p><p><b>RESULTS</b>By using lung cancer cell enrichment and detection kit, it was revealed that the detection rates of CK positive cells were 33.3%, 60.6%, 0% and 0% in Group A, B, C and D, respectively. By using nested RT-PCR, the rates of positive expression of CK19 mRNA were 38.9%, 63.6%, 20.0% and 0% in Group A, B, C and D, respectively, and those of LUNX mRNA were 44.4%, 69.7%, 10.0% and 0%, respectively, while the detection rate of CTCs was negative in all groups using simple ICC. There was a significant difference between the results obtained by lung cancer cell enrichment and detection kit and simple ICC method (P < 0.05), while no significant difference was found between the results obtained by lung cancer cell enrichment and detection kit and nested RT-PCR (P > 0.05). There was a significant difference in the detection rates of CK positive cells between group A and groups B, C, D (P < 0.05). The micrometastasis in peripheral blood was closely related with pathological types, cell differentiation and TNM staging (P < 0.05).</p><p><b>CONCLUSION</b>The lung cancer cell enrichment and detection kit developed by ourselves is a sensitive and reliable method to detect CTCs in patients with NSCLC. It may be helpful in diagnosis of NSCLC micrometastasis and circulating tumor cells in peripheral blood, re-determination of clinical stage and provide important information for cancer-therapy personalization.</p>

Identification of a novel candidate gene KLRC1 within the putative susceptibility locus for systemic lupus erythematosus at 12p12.3-13.2 in a Chinese cohort / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify the candidate genes within the putative susceptibility locus for systemic lupus erythematosus (SLE) at 12p12.3-13.2.</p><p><b>METHODS</b>KLRC1 was selected as the candidate gene according to the results of previous gene chip studies. TaqMan real-time quantitative PCR was performed for detecting KLRC1 mRNA expression in 55 SLE patients and 30 controls.</p><p><b>RESULTS AND CONCLUSION</b>KLRC1 mRNA expression was significantly higher in the mononuclear cells and T cells of SLE patients than in the healthy controls (P<0.01), but showed no significant difference in the B cells. No obvious correlation was found between the SLE disease activity index (SLEDAI) and KLRC1expression level, suggesting that KLRC1 can be a probable candidate gene for SLE on 12p12.3-13.2, but which is not associated with the disease activity.</p>

Retrospective study on 116 unexpected sudden cardiac deaths in Yunnan, China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To identify the epidemiological and clinical features of unexpected sudden cardiac deaths (SUD) in Yunnan.</p><p><b>METHODS</b>Choosing the old SUD cases from Xiangyun, Heqing, Nanjian and Dayao counties and using the standardized verbal autopsy Form, we interviewed the family members of the cases, witnesses and doctors as well as reviewing their medical files to get relative information.</p><p><b>RESULTS</b>We identified 116 SUDs in 21 villages from 1984 to 2004. The village-specific annually standardized incidence rates were ranged from 0.2/1000 to 8.9/1000 (median = 0.8/1000). 66% and 29% of the SUDs occurred in July and August respectively. The incidence rates of SUD were higher (1.6/1000, chi(2) = 16, P < 0.01) in 10 - 39 year-olds, and higher in females than in males (RR = 1.6, 95% CI: 1.1 - 2.3). Seventy percent of SUD occurred in families having clustering nature and 60% of the additional cases in the family were occurred within 24 hours (median = 20 hours) after the first SUD identified in the family. SUD occurred in 23 families followed the first affected family in a village during the same season. In these 23 families, 61% of the first SUD occurred within 8 days after the first SUD in the first affected family. 68% and 66% of the SUDs did not have any complaints or signs during the last 3 weeks or from 3 weeks to 2 days prior to the onset of the disease. 63% of the SUDs had cardiac symptoms within the last 2 days prior to the onset with major symptoms as dizziness, nausea, faintness, unconsciousness, weakness and palpitation. The median duration from acute onset to death was 2 hours.</p><p><b>CONCLUSIONS</b>The extreme time-space clustering of SUD in families and in villages suggested that the risk factors occurred in specific time and location. Familial clustered SUD cases had common exposure pattern. Sudden onset of acute cardiac symptoms often followed by sudden death. Epidemiological study on new cases was necessary to identify risk factors and to develop hypothesis for causation. In July 2005, we instituted a special SUD surveillance system for all the affected counties together with 10 counties which had no reported cases.</p>

Study on a monitoring program regarding leptospirosis in some fore-and-after flood-affected along large rivers in Anhui province / 中华流行病学杂志

<p><b>OBJECTIVE</b>The study was designed to find out the epidemic characteristics of leptospriosis and to develop effective intervention measures. The effects of floods on leptospriosis in some areas along Yangzi river and Huai river in Anhui province was also analysed.</p><p><b>METHODS</b>Study on serum epidemiology of leptospriosis was carried out from serous samples collected from native residents and animal hosts including isolation of pathogens at different phases (before,middle and after) and different monitoring spots,during the floods.</p><p><b>RESULTS</b>Infection rate with leptospriosis pathogen among native residents was 13.49% during the flood-period,much higher than 2.18% at post-flood (chi2 = 22.78, P < 0.01) stage, in the flood-affected areas along Yangzi river in 1998. The average rates of infection were 2.48% and 5.35% in affected and unaffected areas along Huai river respectively, in 2003.</p><p><b>CONCLUSIONS</b>There was full evidence that floods causing the epidemics of leptospriosis. However, the transmission of leptospriosis among people would depend on affecting factors as scales of floods, lasting time, coincidence between flood happening and epidemic season, immuno-protection level against leptospriosis among people and so on to a great extent. Factors as the magnitude of pathogens carried by various kinds of infectious sources were also important determinants affecting the nature, being epidemic or pandemic of leptospriosis. It was suggested that active surveillance network on the sources of infection and risk factors of leptospriosis should be developed for the control and prevention of the disease, in the flood-hit areas.</p>

Application of immunomagnetic screening strategy for separation of CD4+ and CD8+ T cell subpopulations of peripheral blood / 中国实验血液学杂志

To evaluate the separation of T lymphocyte subsets by immunomagnetic beads and to find optimization of strategy for specific binding of antibody-coated beads to cells, two strategies to isolate enriched T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from small volumes (< 5 ml) of peripheral blood by using immunomagnetic beads or complement cytotoxicity method were compared. The purity and activity of CD4+ T cells and CD8+ T cells were measured by using flow cytometry, trypan-blue dye exclusion test, etc. The results showed that the yields of CD4+ T lymphocytes and CD8+ T lymphocytes by using immunomagnetic beads were (94.2 +/- 1.4)% and (93.8 +/- 3.0)% respectively, higher than those of control group and the group of using completement cytotoxicity method (P < 0.05). At the same time, the yields of CD4+ T lymphocytes and CD8+ T lymphocytes by using complement cytotoxicity method were (76.0 +/- 2.8)% and (77.0 +/- 3.0)% respectively, higher than those of unenriched group (P < 0.05). The trypan-blue dye exclusion test confirmed that there were no influences on activity of CD4+ T cells and CD8+ T cells when immunomagnetic beads were used for separation of these cells from peripheral blood. It is concluded that the immunomagnetic bead method has a higher efficiency for separation of CD4+ T cells and CD8+ T cells from peripheral blood than complement cytotoxic method, especially for small sample. This method has no influence on activity and proliferation of T lymphocyte subpopulations, and would be expected to establish conditions for research of biological characteristics of CD4+ T cells and CD8+ T cells in future.

Flow cytometric detection of minimal residual disease in pre-cursor-B-acute lymphoblastic leukemia on the basis of phenotypic aberrancies on minor leukemic cell populations / 中国实验血液学杂志

To test the European BIOMED-1 Concerted Action proposed technique to detect minimal residual disease (MRD) in the chinese patients with precursor-B-acute lymphoblastic leukemia (precursor-B-ALL) by triple-staining flow cytometry and to define both normal and aberrant phenotypic profiles of precursor B cells, a series of bone marrow samples, 35 from precursor-B-ALL (13 in newly diagnosed cases, 15 at the end of remission induction therapy and 7 at end of the consolidations), and 19 from normal controls, were immunophenotyped with the five triple-staining antibodies (TdT/CD10/CD19, CD10/CD20/CD19, CD34/CD38/CD19, CD34/CD22/CD19 and CD19/CD34/CD45) recom-mended by the BIOMED-1 using common flow cytometric protocols. Further, with different ratios of the leukemic cells with CD34/CD38/CD19 phenotype and normal mononuclear cells, a serial dilution test was analyzed. The results showed that three major CD19(+) cell subpopulations were identified in the normal controls, representing three consecutive maturation stages. The subpopulations in the precursor-B-ALL cases disappeared and were replaced with a great number of luekemic cells which had different characteristics of phenotypes, and then they reappeared with almost same characteristics as the normal CD19(+) cells after the patients achieved complete remission. When the five triple-staining antibody combinations were used, the phenotypic aberrancies could be identified in 12/13 (92.3%) cases with newly diagnosed precursor-B-ALL, at least one triple-labeling per case at the level of 0.01% or more. The frequencies of phenotypic aberrations detected with the triple-staining were 8/13 (61.5%) for CD10/CD20/CD19, 5/13 (38.5%) for CD34/CD38/CD19, 4/13 (30.8%) for CD10/TdT/CD19, 3/13 (23.1%) for CD34/CD22/CD19, and 2/13 (15.4%) for CD34/CD45/CD19. At the end of remission induction, the phenotypic aberrancies could be detected in 5/15 (33.3%), of which, 3/8 (37.5%) cases with the leukemic phenotypes detected both at the newly diagnosis and at the end of induction. The dilution test indicated that the cells with CD34/CD38/CD19 detected by flow cytometry correlated well with the leukemic cells added (r = 0.85, P < 0.05) over 1:1 to 1:400,000. It is concluded that the flow cytometric detection of precursor-B-ALL-MRD proposed by BIOMED-1 Concerted Action were well realized in this study. The one precursor-B-ALL cell can be effectively detected out of 10(4) normal bone marrow cells.

The use of bioluminescent image in monitoring islet graft in vivo after islet transplantation / 中华器官移植杂志

Objective To demonstrate the superiority and feasibility of using bioluminescent image to monitor the islet graft after islet transplantation.Methods Diabetic models were established by intraperitoneal injection of streptomycin into mature male C57BL/6 mice.Islets were harvested from the pancreas of C57BL/6 and Bclb/c mice by digestion and purification,and transfected with Lueiferase gene.The mouse diabetic models were divided into iso-transplantaion group (n=20) and allo-transplantation group (n=7).The islets of C57BL/6 were transplanted into iso-transplantaion mice with different doses (10,50,110 and 200,n=5 in every dose),and Bclb/c mouse islets were transplanted into allo-transplantation group.The islets were transplanted into the subcutaneous fat tis- sue near left scapula.The receptor mice were scanned with CCD camera to get bioluminescent images at different scheduled time points,and the changes in random blood glucose of allo-transplantation group were observed.Results On day 6 after transplantation,the scanning image showed that the pi- xel intensity from the region of interest (ROI) was increased with the increased number of islet grafts and they had a positive correlation.The random blood glucose was reduced to the normal level in the first 2 days,and then increased again to the diabetic level on 11 days averagely,while pixel intensity from the ROI reached the peak on day 6-7,and then reduced rapidly after islet transplantation in allo- transplantation group.The beginning of pixel intensity reduction occurred on day (6.14?0.90), while that of the random blood glucose raise occurred on day (10.00?0.82) after transplantation,and the former alteration occurred obviously earlier than the latter (P

Expression and clinical significance of peripheral blood CD4~+CD25~+T cells in patients with systemic lupus erythematosis / 中华风湿病学杂志

Objective To investigate the expression and clinical significance of peripheral blood CD4~+, CD25~+ and CD4~+CD25~+ T subpopulations in patients with systemic lupus erythematosis.Methods The per- centage and fluorescence intensities of peripheral blood CD4~+,CD25~+ and CD4~+CD25~+ subpopulations from 34 SLE and 18 normal controls were measured with flow cytometry assay,then the correlation with clincal data was analyzed.The CD25~+ cells were defined as the CD25~(high) cells if their fluorescence intensity was higher than 10. Results The percentage of CD4~+CD25~+,CD4~+CD25~(high) T lymphocytes in active SLE patients[(4.80?1.21)% and (0.25?0.10)%]was lower than that in normal controls[(8.92?3.21)% and(0.44?0.22)% and non-active SLE patients(11.28?2.09)% and(0.59?0.34)%](P＜0.05).However,as for the CD25~+ cells in the CD4~+ T cells,there was no difference between SLE patients and normal control group.Peripheral blood CD4~+CD25~+,CD4~+CD25~(high) cells in SLE were reversely correlated with SLEDAI(r=-0.74,P=0.004 and r=-0.614,P=0.026),but not with others such as complements,ANA titers etc.Peripheral blood CD4~+ and CD25~+ lymphocytes in active SLE pa- tients were also lower than those in normal controls[(23?7)vs(34?7)and(7.4?1.8)vs(13.9?3.4),P＜0.05]. CD25 fluorescence intensities were higher in the SLE patients those in the normal controls,but CD4 fluores- cence intensities were not.Conclusion CD4~+CD25~+ may play a role in the pathogenesis of SLE.

Acquired hemophilia A in patients with systemic lupus erythematosus:report of a case and review of literature / 中华风湿病学杂志

Objective To summarize the manifestation and treatment of acquired hemophilia A in pa- tients with systemic lupus erythematosus(SLE).Method A case was investigated retrospectively and the lit- erature was reviewed.Results A 25-year-old woman with a 5 year history of SLE was admitted to hospital due to abdominal pain.She was diagnosed with acquired factorⅧinhibitor deficiency based on a prolonged activated partial-thromboplastin time(APTT,135.3 s),reduced factorⅧactivity(0.9%)and factorⅧin- hibitor(26.1 BU/ml).Sonography and magnetic nuclear resonance of the abdomen confirmed the presence of a retro-uterine hematoma.The patient was initially treated with intravenous pulse and oral corticosteroids,factorⅧplasma concentrated and intravenous immunoglobulin.Clinical and biological improvement was promptly obtained.Conclusions Attention should be paid to the association between SLE and acquired hemophilia A. Combination therapy may be recommended as initial therapy for the management of acquired hemophilia A in patients with SLE.But no standardized treatment can be recommended at present.