Establishment of Flow Cytometric Immunobead Array Assay for Quantitation of Platelet-Specific Antibodies and Its Application / 中国实验血液学杂志

OBJECTIVE@#To establish a flow cytometric immunobead array assay (FCIA) to quantify platelet antibodies and to explore its application in the diagnosis and treatment of ITP.@*METHODS@#The guantitative standard curve was established by binding the human IgG of known concentration on antibody-coated microbeads; at the same time, the platelet-specific antigen and antibody complex was captured and levels of platelet antibodies were detected using the microbeads coated by 5 kinds of antibodies against platelets suca as GPIX (SZ1), GPⅠb (SZ2), GpⅢa (SZ21), GPⅡb (SZ22) and p-selection (SZ51). The fluorescence signal detected by flow cytometry were transformed into the conentration of platelet antibodies in samples through the quantitative standard curve, thereby establishing the method for quantititive detection of platelet-specific antibodies in plasm samples (FCIA), moreover the property, efficiency and clinical application of establishod FCIA method were evaluated.@*RESULTS@#The FCIA could detect 5 kinds of antibodies against GPIX, GPⅠb, GpⅢa, GPⅡb and β-selection within a broad range of 33.29-1280 ng/ml, 45.17-1280 ng/ml, 42.07-1280 ng/ml, 46.40-1280 ng/ml, 42.48-1280 ng/ml and 42.48-1280 ng/ml respectively, and their recovery rates were 115.23%, 112.58%, 117.47%, 107.64% and 112.67% respectively. The intra-assay coefficient of variation (CV) for anti- GPIX, -GPⅠb, -GpⅢa, -GPⅡb and p-selection antibodies was 3.54%, 3.63%, 4.66%, 6.43% and 6.67% respectively, and the inter-assay CV for above mentioned antibodies were 10.89%, 7.57%, 10.34%, 6.95% and 10.72% respectively. The detection showed that the levels of 5 kinds of platelet-specific antibodies in ITP group all were higher than those in non-ITP and healthy control groups (P＜0.01). The sensitivity, specificity and accuracy of quantitatively detecting 5 kinds of antibodies for diagnosis of ITP by FCIA were 68.29%, 84.98% and 78.95% respectively, while the sensitivity, specificity and accuracy of detecting 5 kinds of antibodies by modified indirect MAIPA were 41.46%, 90.41% and 72.81% respectively.@*CONCLUSION@#The established quantitative FCIA for detection of antibodies provides a powerful tool for diaghosis and evaluation of therapeutic efficacy and prognosis of ITP patients.

Cathelicidin LL-37 restoring glucocorticoid function in smoking and lipopolysaccharide-induced airway inflammation in rats / 中华医学杂志(英文版)

BACKGROUND@#Glucocorticoids have been widely used to treat patients with chronic obstructive pulmonary disease (COPD). Nevertheless, corticosteroid insensitivity is a major barrier to the effective treatment of COPD and its mechanism remains unclear. This study aimed to evaluate the effect of cathelicidin LL-37 on corticosteroid insensitivity in COPD rat model, and to explore the involved mechanisms.@*METHODS@#COPD model was established by exposing male Wistar rats to cigarette smoke combined with intratracheal instillation of lipopolysaccharide (LPS). Inhaled budesonide and LL-37 were consequently applied to COPD models separately or collectively to confirm the effects on inflammatory cytokines (tumor necrosis factor [TNF]-α and transforming growth factor [TGF]-β) by enzyme-linked immunosorbent assay (ELISA) and lung tissue histopathological morphology. Expression of histone deacetylase-2 (HDAC2) and phosphorylation of Akt (p-AKT) in lung were also measured.@*RESULTS@#Briefly, COPD model rats showed an increased basal release of inflammatory cytokines (lung TNF-α: 45.7 ± 6.1 vs. 20.1 ± 3.8 pg/mL, P < 0.01; serum TNF-α: 8.9 ± 1.2 vs. 6.7 ± 0.5 pg/mL, P = 0.01; lung TGF-β: 122.4 ± 20.8 vs. 81.9 ± 10.8 pg/mL, P < 0.01; serum TGF-β: 38.9 ± 8.5 vs. 20.6 ± 2.3 pg/mL, P < 0.01) and COPD related lung tissue histopathological changes, as well as corticosteroid resistance molecular profile characterized by an increase in phosphoinositide 3-kinase (PI3K)/Akt (0.5 ± 0.1 fold of control vs. 0.2 ± 0.1 fold of control, P = 0.04) and a decrease in HDAC2 expression and activity (expression: 13.1 ± 0.4 μmol/μg vs. 17.4 ± 1.1 μmol/μg, P < 0.01; activity: 1.1 ± 0.1 unit vs. 1.4 ± 0.1 unit, P < 0.01), compared with control group. In addition, LL-37 enhanced the anti-inflammatory effect of budesonide in an additive manner. Treatment with combination of inhaled corticosteroids (ICS) and LL-37 led to a significant increase of HDAC2 expression and activity (expression: 15.7 ± 0.4 μmol/μg vs. 14.1 ± 0.9 μmol/μg, P < 0.01; activity: 1.3 ± 0.1 unit vs. 1.0 ± 0.1 unit, P < 0.01), along with decrease of p-AKT compared to budesonide monotherapy (0.1 ± 0.0 fold of control vs. 0.3 ± 0.1 fold of control, P < 0.01).@*CONCLUSIONS@#This study suggested that LL-37 could improve the anti-inflammatory activity of budesonide in cigarette smoke and LPS-induced COPD rat model by enhancing the expression and activity of HDAC2. The mechanism of this function of LL-37 might involve the inhibition of PI3K/Akt pathway.

Application of CRISPR-Cas9 technology in non-cancerous hematological disorders / 上海交通大学学报（医学版）

The recent discovery of the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 9 (Cas9) system for precise genome editing has revolutionized methodologies in hematology studies. CRISPR-Cas9 technology can remove and correct genes or mutations, and introduce site-specific therapeutic genes in human cells. Therefore, it has become the ideal targets for inherited haematological disorders and could be the choice to alleviate disease-related symptoms by correcting disease-causing mutations in the near future. Prior to the use of CRISPR-Cas9-mediated gene correction in humans, appropriate delivery systems with high efficiency and specificity must be identified, and ethical guidelines for applying the technology with controllable safety must be established. In this review, the latest applications of CRISPR-Cas9 technology in haematological disorders, current challenges and future directions were discussed.

Analysis of NK Cells and Receptors in Peripheral Blood of Patients with HBV Related Acute-on-chronic Liver Failure in Different Phases / 中山大学学报(医学科学版)

[Objective]To investigate the character of expression of NK cells and its receptors in peripheral blood of pa-tients with HBV-related acute-on-chronic liver failure(HBV-ACLF)in different phase.[Methods]There are thirteen pa-tients with HBV-ACLF in advanced phase group and thirty-three patients in plateau phase group,with thirteen healthy per-son in control group.The frequency of NK cells and expression of NK cell activating receptors(NKAR)including NKG2D, NKp30 and NKp46,NK cell inhibitory receptors(NKIR))cludingsd ate the character of including NKG2A,KIR2DL1, KIR2DL3 and KIR3DL1,NK cell killing function related factors including Perforin,GranzymeB and FasL in peripheral blood of all patients were detected by flow cytometry.[Results]The percentage of NK cells in advanced group was lower than plateau group and healthy control group(H=7.771,P=0.021).The expression of KIR3DL1 in healthy control group was high-er than advanced group and plateau group(Z=6.639,P=0.036)while the expression of FasL was significantly lower than ad-vanced group and plateau group(Z=22.5,P<0.001).[Conclusion]Patients in advanced group had lower frequency of NK cells,lower expression of inhibitory receptor KIR3DL1 and higher expression of FasL than patients in plateau group and healthy control group,which is associated with immune status of patients in different phase of liver failure.

Comparison of embryonic nature orifice transumbilical endoscopic surgery with conventional laparoscopic surgery treatmenting Hirschsprung disease / 中华实用儿科临床杂志

Objective To compare the therapeutic effects of embryonic nature orifice transumbilical endoscopic surgery(ENOTES) with conventional laparoscopic(CL) surgery in Hirschsprung disease(HD).Methods A total of 111 children from Sep.2009 to Dec.2010 were included in the retrospective study.The patients were divided into 2groups:ENOTES group (66 cases) and CL group (45 cases).The general conditions of children were reviewed operative situation,postoperative complications and defecation 1 year after operation.Results In the left colectomy,there was no difference between 2 groups in age,weight,operative blood loss and postoperative stay (all P ＞ 0.05),but ENOTES took up less time than CL(P ＜ 0.05) ;in the subtotal colectomy,no significant difference existed in all series.All children underwent primary radical surgery,neither transfer to open surgery nor dead case.In ENOTES group,there were 9 cases suffering from enterocolitis postoperatively (13.6％),while 7 cases (15.6％) in CL group.Twenty-eight patients in ENOTES group and 12 in CL group were followed up for 1 year.Less abdominal distension postoperatively was noted in ENTOES group than CL group(P ＜ 0.05),but no other difference in other available data.In addition,no obvious operative scar was noted on the abdomen in ENOTES group,what meant better cosmetic benefit than CL.Conclusion HD treated with ENOTES can gain the same outcomes with CL with better cosmetic effect.

Transumbilical laparoscopic pull-through for children with hypoganglionosis / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the surgical outcomes after on transumbilical laparoscopic pull-through procedure for pediatric hypoganglionosis(HYP).</p><p><b>METHODS</b>Twelve children with HYP had received transumbilical laparoscopic pull-through procedure from June 2009 to June 2010. Specially designed curved and elongated laparoscopic instruments were used during the procedures. All the patients were followed up over 10 months. Data were collected and analyzed. The diagnosis of hypoganglionsis was pathologically confirmed.</p><p><b>RESULTS</b>No conversions to laparotomy or traditional laparoscopic surgery were required and there were no damages to the abdominal blood vessels, intestine, ductus deferens, or ureters. The average duration of operation was 140 min. The mean intraoperative blood loss was 45 ml. The mean length of specimen was 40 cm. Postoperatively there were no complications such as anastomotic leak, anastomotic stricture, constipation, seepage, or fecal in continence. The average hospital stay after surgery was 9 days. During 10 to 22 months of follow-up(median 16 months), no postoperative recurrence was noticed. No obvious scar was seen 1 months after surgery.</p><p><b>CONCLUSION</b>It is safe and effective for children with hypoganglionosis to undergo transumbilical laparoscopic pull-through procedure.</p>

Effects of allogenic intra-bone marrow bone marrow transplantation on the hematopoiesis in mice / 基础医学与临床

Objective To investigate the effects of allogenic intra-bone marrow bone marrow transplantation (IBM-BMT) on re-establishing hematopoiesis in mice. Methods Bone marrow mononuclear cells (BMNCs) from BALB/ c mice were transplanted into the C57BL/6 mice treated with a lethal dose of ~(60)Coγ-ray radiation through intra-bone marrow injection or intravenous injection. Sixty of the C57BL/6 mice were randomly divided into three groups as higher dose intra-bone marrow injection group (IBM1 group), lower dose intra-bone marrow injection group (IBM2 group) and intravenous injection group (IV group). The nucleated cell numbers of whole bone marrow from the tibia of each recipient mouse were counted respectively at the day 1, day 3, day 6 and day 9 after the transplantation. The donor-derived total nucleated cells and myeloid cells were quantified by flow cytometry. Results At 6th day after transplantation, more total bone marrow nucleated cells, total donor-derived nucleated cells and donor-derived myeloid cells in the tibia of injected side in both IBM1 group and IBM2 group were found than that in IV group (P<0.05 or P<0.01). Conclusion Compared with traditional bone marrow transplantation (IV-BMT),IBM-BMT improves the bone marrow hematopoiesis in the early hematopoietic re-establishing stage in allogenic bone marrow transplantation.

Reoperation for postoperative constipation recurrences of Hirschsprung disease / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To summarize the causes and treatment experiences of postoperative constipation recurrences of Hirschsprung disease.</p><p><b>METHODS</b>Clinical data of 37 cases receiving operation again for postoperative constipation recurrences of Hirschsprung disease were collected. The recurrent causes, reoperation procedures and therapeutic efficacy were analyzed.</p><p><b>RESULTS</b>Among 37 cases, the recurrent causes included insufficient resection of the intestine in 14 cases, co-existent intestinal neural hypogenesis in 6, gate syndrome in 5, and others in 12 cases. There was no postoperative death. Thirty-one cases (83.8%) were followed-up from 6 months to 11 years. The defecation frequency was 1-2 times per day in 15 cases, one times per two days in 7, one times per three days in 6 cases. Two cases could defecate with the help of laxative purgatives. Nobody had incontinence.</p><p><b>CONCLUSION</b>The main cause of postoperative constipation recurrences of Hirschsprung disease is insufficient resection of the intestine,and reoperation can get better results.</p>

Behavior Therapy for relieving the symptoms of outpatients with schizophrenia / 中国康复理论与实践

@#ObjectiveTo study the effects of the Behavior Therapy on outpatient with schizophrenics in relieving symptoms, insight, and attitude to treatment. Methods96 outpatients with schizophrenia were randomly assigned to the Behavior Therapy group and the control group. The Behavior Therapy courses were given to the Behavior Therapy group for six months. All the subjects were evaluated with Positive and Negative Syndrome Scale(PANSS) and Insight and Therapeutic Attitude questionaire(ITAQ). Results94 cases completed the trial. The scores of PANSS, the Negative Subscale, the General Psychopathology Subscale of the Behavior Therapy group decreased significantly after treatment(P<0.01). The score of ITAQ of the Behavior Therapy group increased significantly after treatment(P<0.01).The reductions of PANSS, the Positive Subscale, the Negative Subscale, the General Psychopathology Subscale and ITAQ between two groups were significant different(P<0.05-P<0.01). Conclusions The Behavior Therapy may play an important role in augmenting optimal pharmacotherapy for outpatients with schizophrenia and it is effective in improving symptoms, insight and therapeutic attitude.

A follow-up study of effect of the two kinds of skills training for rehabilitating schizophrenics in the community / 中国康复理论与实践

@#ObjectiveTo explore the efficacy of medication management and symptom management skills-training for preventing schizophrenics' relapse and rehabilitating their mental handicaps.Methods133 subjects were randomly assigned to the skills training group and the control group. Both groups received the same treatment, but the skills training courses were given to the skills training group for the first twenty weeks. One-year follow-up was carried out. All subjects were evaluated with standard rating scales and self-complied drug treatment compliance rating scale. Results128 subjects had completed the research. The skills training group demonstrated clinical results significantly superior to those of the control group on overall improvement according to the total score of the drug treatment compliance rating scale, the rate of relapse, the rate of re-hospitalization and the rate of effectiveness of minimizing handicap(147.9±53.2 vs. 90.4±16.3, 146.1±20.0 vs. 91.7±16.7;12.5% vs. 55%; 3.2% vs. 39.6%;86.5% vs. 26.5%, P<0.01).Conclusions The two kinds of skills training are effective in both preventing the relapse of schizophrenics in the community and minimizing their handicap.