With the advantage of being capable of detecting multiple targets at the same time, high throughput and cost-effective, multiplex nucleic acid detection technologies meet the need of large-scale nucleic acid screening and quantification. Multiplex polymerase chain reaction has been applied to detect pathogen, methylated DNA, mutated gene, and single nucleotide polymorphism typing. Isothermal amplification technologies, such as loop-mediated isothermal amplification and recombinase polymerase amplification are promising in the field of point-of-care testing. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas)-based multiplex nucleic acid detection technologies have become a hotspot due to their high sensitivity and specificity. Metagenomics sequencing plays a leading role in the detection of emerging pathogens and their gene mutation monitoring as well as tumor research. In this review, the advancements and future of multiplex acid detection technologies in clinical application are discussed.

OBJECTIVETo analyze the clinical and molecular characteristics of patients with X-linked thrombocytopenia (XLT) and their responsiveness to treatment with various doses of corticosteroids or intravenous immunoglobulin (IVIG) separately.

METHODData from 15 XLT patients who were hospitalized in Children's Hospital Affiliated to Chongqing Medical University from March 2010 to July 2014 were analyzed retrospectively, including clinical manifestations, scores, peripheral blood, immunological functions, responses to IVIG and steroid treatment with various doses and duration.

RESULTAll 15 XLT patients met the inclusion criteria and showed microthrombocytopenia with or without mild-to-moderate eczema or minor infections. Platelet counts ranged from (8-80) × 10⁹/L. The platelet volume value ranged between 5.6 and 10.9 fl (normal range: 9.4-12.5 fl). Raised serum IgG was found in 5 cases, while low serum IgG was found in 2 cases. WAS gene analysis revealed missense mutations in 14 patients, including 4 hotspots (V75M, R86C, R86H, R86L) and 1 novel mutation (Y107C). Flow cytometer analysis of 13 patients showed various amounts of WAS protein (WASP) expression, 2 patients had normal amounts of WASP expression, 5 had reduced amounts, and 6 had absent WASP expression. Their responses to individual steroid and IVIG treatment with various doses and duration were also reviewed. Fourteen patients who were misdiagnosed as immune thrombocytopenic purpura at first received 28 courses of steroids and (or) 47 courses of IVIG treatment. The post-treatment platelet counts of 1 000-2 000 mg/(kg × d) IVIG(25 courses) at 2-7 d and 8-14 d time points were (60 ± 10) × 10⁹/L and (41 ± 7) × 10⁹/L, which indicate a significantly better responsiveness than those by [(31 ± 7) × 10⁹/L, (21 ± 2) × 10⁹/L] of 400-500 mg/(kg·d) IVIG(22 courses) (Z = -4.419, -1.592;P = 0.002,0.011). However, there were no significant differences between the responsiveness of 3 doses [1-2 mg/(kg·d)(8 courses), 3-6 mg/(kg·d) (11 courses) and 20-30 mg/(kg × d)(9 courses)] of steroids (F = 0.387,0.252;P = 0.980,0.761) at 2-7 d and 8-14 d time points. The platelet counts gradually decreased to the primary level at 15-30 d after any doses of steroids and (or) IVIG treatment. The effective rate of 1 000-2 000 mg/(kg × d) IVIG treatment was 18/25, which was significantly higher than that (2/22) of 400-500 mg/(kg × d) (χ² = 9.836, P = 0.008). The effective rate of 20-30 mg/(kg × d) steroids treatment (7/9) was relatively higher than 1-2 mg/(kg × d) (4/8) and 3-6 mg/(kg × d) (6/11) with no significant difference (χ⁹ = 3.235, P = 0.581). After the treatment with steroids and /or IVIG 14 cases with hemorrhage were all improved.

CONCLUSIONThe clinical characteristics of X-linked thrombocytopenia were microthrombocytopenia with or without mild-to-moderate eczema or minor infections. WAS gene and WASP analysis were diagnostic methods. There were no significant differences between the responsiveness of 3 doses of steroids; 1 000-2 000 mg/(kg·d) IVIG had a significantly better responsiveness. However, IVIG and steroids with any dose and duration may only transiently increase peripheral platelet level of XLT patients.

Adrenal Cortex Hormones ; administration & dosage ; Child ; Genetic Diseases, X-Linked ; drug therapy ; Humans ; Immunoglobulins, Intravenous ; administration & dosage ; Platelet Count ; statistics & numerical data ; Retrospective Studies ; Thrombocytopenia ; drug therapy ; Treatment Outcome

ObjectiveUsing dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) to evaluate the hemodynamic perfusion characteristics of bone marrow infiltration in patients with acute leukemia (AL). MethodsForty-seven patients with AL received coronal pelvic T1WI DCE-MRI with fast low angle shot (FLASH) sequence. Among them, 25 were initial onset untreated (IOU) patients, 22 were treated AL patients, including 14 with complete remission (CR) and 8 with non-remission ( NR). The hemodynamic perfusion parameters including maximum percentage of enhancement ( Emax ) and slope were determined based on enhancement-time curves ( ETCs ) of iliac and lumbar vertebra. The proportion of marrow myeloblasts was recorded.For all patients, quantitative perfusion parameters of bone marrow infiltration in ilium were compared with those in lumbar. The values of Emax and ES were compared among IOU,CR and NR patients.Correlations between perfusion parameters and histopathological results were assessed. ResultsIn all the 47 patients, the Emax values of bilateral iliac bone marrow ( 15.70 ± 7.06)were slightly higher than that of lumbar bone marrow ( 11. 28 ± 5.52 ), and the difference was statistically significant (P ＜0. 01 ).There was no significant difference in the slop value between bilateral iliac bone marrow (0. 82 ± 0. 12 ) and lumbar bone marrow (0. 80 ± 0. 09 ) ( P ＞ 0. 05 ). In the 25 untreated patients,the Emax and slop values were 17. 15 ± 5.75 and 0. 98 ± 0. 13, respectively; in the 14 CR patients, they were 8. 76 ±3.93 and 0. 26 ± 0. 04, respectively, and in the 8 NR patients, they were 21.62 ± 6. 50 and 1. 38 ± 0. 02, respectively. There was significant difference in the Emax and slop values among the three groups (P＜0. 05).Compared with IOU and NR patients, both the Emax and slop values decreased significantly in iliac bone marrow of AL patients with CR (P ＜ 0. 05 ). There was no significant difference between IOU and NR patients ( P ＞ 0. 05 ). A significant positive correlation was found between Emax value of iliac bone marrow and the proportion of marrow myeloblasts ( r ＝0. 501 ,P ＜0. 05 ). There was a negative correlation between slop value of iliac bone marrow and the proportion of marrow myeloblasts ( r ＝0. 235 ,P ＞0.05).ConclusionsDCE-MRI can beused for evaluating the hemedynamic characteristics of microcirculation of bone marrow infiltration in patients with AL, which can provide useful information in evaluating prognosis and monitoring therapeutic effect.

ObjectiveTo report the preliminary results and surgical technique of Stoppa approach used in treatment of acetabular fracture.MethodsFrom May 2005 to May 2010,23 patients with acetabular fracture were treated using modified Stoppa approach.The mean age was 37 years old(range,21-71years old).According to Letournel classification,there were 7 cases in anterior column fracture,6 cases in anterior column with posterior hemitransverse fracture,6 cases in transverse fracture,4 cases in associated with both column fractures.All patients in the study were operated under general anesthesia on a radiolucent table in a supine position and underwent Stoppa approach.Fracture reduction was evaluated by Matta's score system.The clinical outcome was judged by modified Merle d' Aubigne and Postel score system.Four cases of associated both column fractures,4 cases of anterior column with posterior hemitransverse fracture required the use of the lateral window.ResultsThe mean surgical time was 166 min (range,110-320 min).The mean blood lose was 647 ml (range,300-2500 ml).Blood transfusion was 3.3 unites (0-12 unites).All the factures healed in 3 months.The fracture reduction was evaluated according to the Matta's score system:an excellent reduction was obtained in 15 patients(65％),a good reduction was obtained in 5 patients (22％),and three(13％) were considered poor.Clinical results were excellent in 11 patients(48％),good in 7 patients (30％),fair in 3 patient (13％),and poor in 2 patients (9％).Superficial wound infection happened in 1 case.It healed after debridment.One patient noted to have significant weakness of the hip adductors (obturator nerve palsy) after surgery.It reached grade Ⅳone year later.No sciatic nerve,femoral nerve and vascular injury happened in this study.ConclusionFor the selected acetabualar fracture,Stoppa approach can get satisfied exposure and fixation of the fracture,especially in the patient with central dislocation of the femoral head as well as medial displacement of quadrilateral plate.

[Objective] To investigate the effects of Yangyin Kangdu Powder (YKP) on peripheral blood T-lymphocyte subsets levels in X-ray-irradiated mice. [Methods] Forty-five mice were randomized into normal control, model and YKP groups. Except the normal group, the rats in model and YKP groups were irradiated with 6Gy X-ray to establish models with acute radiation-induced injury. Normal saline was given to the model group while YKP in the dosage of 10 g?kg-1?d-1 was orally administered to YKP group for one week. The changes of CD4 and CD8 T-lymphocyte subsets levels were detected by flow cytometry. [Results] Plasma levels of CD4 and CD8 and the ratio of CD4/CD8 decreased in the model group, the difference being significant as compared with the normal control group (P

Objective To investigate the effect of glucocorticoid on ischemia-induced neuronal damage in gerbil hippocampus with forebrain ischemia model.Methods Twenty-one gerbils were randomly assigned to 3 groups (7 animals each). The animal was anesthetized and both common carotid arteries were exposed and separated. Silk threads were looped around these arteries. In group A and group B ,10 ?l saline was given and in group C dexamethasone-water soluble ( 3?g dissolved in 10 ?l saline ) was administered intracerebroventricularly. After a stabilization period of 60 min, transient forebrain ishcmia for 2.5 min was induced in group B and group C by pulling the arteries with 8g weights under the brain temperature of 37.5℃?0.2℃. Seven days after the ischemia, the brains were taken out and fixed with 10% buffered formalin. Brain slices, 5 ?m thick, were stained with hematoxylin and eosin. The numbers of preserved pyramidal cells in the hippocampal CA1 field per 1 mm length of stratum pyramidal were counted.Results (1) Compared with those in group A, the preserved pyramidal cells in hippocampal CA1 field were reduced in group B and group C (P