Objective To investigate the application of fetuses with talipes equinovarus (TE) using chromosomal microarray analysis (CMA) technology. Methods From May 2012 to June 2015, 54 fetuses were found with TE and with or without other structural anomalies by prenatal ultrasound. Karyotyping was taking for them all, and the fetuses with normal karyotypes took another CMA test. The data were analyzed with CHAS software. Finally all the cases were followed up to know about their pregnancy outcomes. Results One of the 54 cases was detected with abnormal karyotype which was trisomy 18 (2%, 1/54). CMA was undertaken to the remaining fetuses, they were divided into 2 groups, including isolated TE group (n=38) and complex TE group (n=15). The detection rate of clinical significant copy number variations (CNV) by CMA was 11% (6/53), while isolated and complex TE group were 5% (2/38) and 4/15, respectively (P=0.047). Of the 53 cases, 51 cases were successfully followed up. Eleven cases were found without TE after birth, and the false positive rate (FPR) of TE was 22%(11/51). Conclusions Whole-genome high-resolution CMA increased the detection rate by 11% in fetuses with TE. With the FPR and the detection rate of the clinical significant CNV of 2 groups, whole-genome CMA could be recommended to the fetuses with complex TE group but normal karyotypes. A series of ultrasonic tests should be suggested to the isolate TE group, while with the abnormal ultrasound, fetuses would be suggested to have CMA test for decreasing the rates of invasive prenatal diagnosis and FPR.

AIMTo study the pharmacokinetic (PK) properties in rabbits treated with N-Ile(1)-Thr(2)-63-desulfato-r-hirudin (rH) newly developed in China by means of bioassay in order to provide preclinical experiment basis for its development as a novel anticoagulant agent.

METHODSrH plasma concentration was determined using bioassay based on ex vivo antithrombin activity of rH. Normal rabbits received iv rH 4.0, 2.0 and 1.0 mg/kg or sc rH 2.0 mg/kg, respectively. The rabbits with acute severe renal failure were given iv rH 2.0 mg/kg.

RESULTSThe bioassay described in this paper met requirements for study of PK in rabbits. The major PK parameters after iv dosing were as follows: t(1/2beta) 58.4-59 min. V(d) 0.09-0.12 L/kg, CL 0.0035-0.0040 L/(kg.min); AUC were proportional to the doses, t(1/2) and CL did not change significantly with the doses. The sc bioavailability reached 94%. The rabbits suffering from acute severe renal failure presented 11-fold longer t(1/2beta) and 13-fold greater AUC than normal healthy rabbits.

CONCLUSIONrH exhibited rapid elimination, distribution was only limited to extracellular space and good absorption from sc site. The excretion of rH by kidneys played a very important role in the elimination of rH. The PK of rH could be described by the two- and one-compartment model after iv and sc dosing, respectively, and followed linear kinetics.

Algorithms ; Animals ; Biological Assay ; methods ; Computer Simulation ; Hirudins ; blood ; pharmacokinetics ; Metabolic Clearance Rate ; Models, Biological ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity ; Thrombin Time ; methods

OBJECTIVETo establish an artificial somatic-autonomic reflex arc in rats and observe the following distributive changes of neural fibers in the bladder.

METHODSAdult Sprague-Dawley rats were randomly divided into three groups: control group, spinal cord injury (SCI) group, and reinnervation group. DiI retrograde tracing was used to verify establishment of the model and to investigate the transport function of the regenerated efferent axons in the new reflex arc. Choline acetyltransferase (ChAT) in the DiI-labeled neurons was detected by immunohistochemistry. Distribution of neural fibers in the bladder was observed by acetylcholine esterase staining.

RESULTSDiI-labeled neurons distributed mainly in the left ventral horn from L3 to L5, and some of them were also ChAT-positive. The neural fibers in the bladder detrusor reduced remarkably in the SCI group compared with the control (P < 0.05). After establishment of the somatic-autonomic reflex arc in the reinnervation group, the number of ipsilateral fibers in the bladder increased markedly compared with the SCI group (P < 0.05), though still much less than that in the control (P < 0.05).

CONCLUSIONThe efferent branches of the somatic nerves may grow and replace the parasympathetic preganglionic axons through axonal regeneration. Acetylcholine is still the major neurotransmitter of the new reflex arc. The controllability of detrusor may be promoted when it is reinnervated by the pelvic ganglia efferent somatic motor fibers from the postganglionic axons.

Acetylcholinesterase ; biosynthesis ; Anastomosis, Surgical ; Animals ; Autonomic Fibers, Preganglionic ; physiology ; Cholinergic Fibers ; metabolism ; Immunohistochemistry ; Motor Neurons ; cytology ; metabolism ; Nerve Regeneration ; physiology ; Neural Pathways ; cytology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reflex ; physiology ; Spinal Cord Injuries ; physiopathology ; Spinal Nerve Roots ; surgery ; Urinary Bladder ; innervation ; physiology ; surgery ; Urinary Bladder, Neurogenic ; surgery

OBJECTIVETo evaluate the therapeutic effect of combined reconstruction of anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) simultaneously by using allograft patellar tendon under arthroscopy.

METHODSFrom May 2003 to November 2005, 10 cases of ruptured ACL and PCL were fixated with compressed screws and reconstructed under arthroscopy with allograft patellar tendon simultaneously. The clinical results were evaluated according to IKDC, Lysholm, and Tegner clinical rating scales.

RESULTSAll patients were followed up for 12-30 months (mean: 18 months). At the last follow-up, there was no knee extension limitation and knee flexion was between 120 degree and 135 degree,with an average of 128.38 degree. The Lysholm score of the 10 cases was 66.5+/-5.6 before operation and 89.8+/-3.4 at last follow up. The difference was statistically significant (P less than 0.01). The average Tegner activity score decreased from 6.9+/-1.7 (range: 4-9) before injury to 5.5+/-1.6 (rang:2-9) at the follow-up (P equal to 0.53). At the end of follow-up, IKDC score was graded as A in 4 cases (40.0%), B in 5 (50.0%), and C in 1 (10.0%). Of the 10 patients, 8 returned to the same sports level as before injury and 2 were under the level.

CONCLUSIONArthroscopic combined reconstruction of ACL and PCL with allograft patellar tendon has the advantages of minimal trauma in surgery and reliable satisfactory outcome.

Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Arthroscopy ; Female ; Follow-Up Studies ; Humans ; Male ; Patellar Ligament ; transplantation ; Posterior Cruciate Ligament ; injuries ; surgery ; Reconstructive Surgical Procedures ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo evaluate the clinical result of arthroscopic anterior cruciate ligament (ACL) reconstruction with either allo- or auto- bone-patellar tendon-bone (B-PT-B) grafts.

METHODSFrom February 2002 to January 2006, 142 of 187 cases of ACL ruptures who received ACL reconstruction with B-PT-B grafts were studied retrospectively. There were 93 male and 49 female whose age was from 15 to 57 years (mean 26 years). Patients were divided into 2 groups by graft selection: 38 patients with autograft and 104 with allograft. Clinical results were evaluated according to IKDC, Lysholm, Irgang and Larson scales. Sub-items of scales such as pain, swelling and laxity were specifically evaluated.

RESULTSAll of the patients were followed up with an average of 24 months (range from 6 to 43 months). All grafts were radiographically in good position at the time of follow-up. KT-1000 examination of affected knee showed less than 3 mm anterior translation difference compared with contralateral one's. Allograft group: 85 patients got normal IKDC score (81.7%). Lysholm score 82.8 +/- 8.5, Irgang score 79.2 +/- 7.3, Larson score 86.7 +/- 3.1. Autograft group: 29 patients got normal IKDC score (76.3%). Lysholm score 84.6 +/- 9.5, Irgang score 79.5 +/- 7.6, Larson score 88.9 +/- 6.8. No significant statistical difference was found between the 2 groups in the comprehend scale (P>0.05).

CONCLUSIONSBoth autograft and allograft group achieve good results, and the allograft B-PT-B could provide the similar clinical result as autograft, the preliminary result of allograft reconstruction might indicate predictable result in the future.

Adolescent ; Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Arthroscopy ; Bone-Patellar Tendon-Bone Grafting ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Matched-Pair Analysis ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Transplantation, Autologous ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo analyze patients with skeletal anomalies (SA) but a normal karyotype using chromosome microarray analysis (CMA).

METHODSFrom June 2012 to May 2015, 43 children found to have skeletal anomalies with or without other abnormalities were subjected to karyotyping analysis. For those with a normal karyotype, DNA was extracted and hybridized with Affymetrix CytoScan 750 kb arrays following the manufacturer's protocol. The results were analyzed with CHAS v2.0 software.

RESULTSTwo patients (4.65%) were detected with an abnormal karyotype. The remaining 41 patients with a normal karyotype were classified into 3 groups: isolated SA (n=17), SA with mental retardation (n=6), and SA with other structural anomalies (n=18). Clinically significant copy number variations (CNVs) were found in 21.95% (9/41) of the cases, which included 17.65% (3/17) with isolated SA, 33.33% (2/6) with SA and mental retardation, and 22.22% (4/18) of SA with other structural deformities.

CONCLUSIONWhole-genome CMA can detect clinically significant CNVs which may not be found by conventional karyotyping analysis and increase the detection rate by approximately 21.95%. It may be recommended for patients with SA but a normal karyotype.

Bone and Bones ; abnormalities ; Child ; Child, Preschool ; Chromosome Aberrations ; DNA Copy Number Variations ; Humans ; Infant ; Infant, Newborn ; Karyotype ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo observe degenerative intervertebral disc and to examine innervation of degenerative discs in the rabbit anular-injury model.

METHODSTwo different magnitudes of anular injury at 5 mm depth were performed by 11 blade or 16 gauge needle at the L3-L4 or L5-L6 discs in New Zealand white rabbits (n=48, 2.5-3.0 kg). Disc degeneration was evaluated by radiographic, MRI and histological examination at different time points after surgery. To identify nerve ingrowth into disc, two general markers PGP 9.5 and GAP 43, for nerve fibers were examined by immunohistochemistry.

RESULTSignificant decreases in disc height and signal intensity in magnetic resonance imaging were observed in 11 blade group and 16 G puncture group (P<0.01). 16 G puncture group induced slower and more progressive disc degeneration companed with the stab group and control group. At the 12-week time point, nucleus pulposus tissues were extruded and scar tissues formed outside the disc. In stab discs, nerve ingrowth was scattered on the surface of injury site and in the deeper part of the scar tissues, more than 1 mm from the surface. However, in punctured discs, PGP 9.5 and GAP 43-immunoreative fibers were only observed in the outmost part of the scar tissues and superficial area. More nerve fibers were observed in stab group.

CONCLUSIONInnervation may act as a source of discogenic pain which is associated with intervertebral disc degeneration caused by disc anular injury.

Animals ; GAP-43 Protein ; metabolism ; Intervertebral Disc ; injuries ; innervation ; pathology ; Intervertebral Disc Degeneration ; diagnosis ; diagnostic imaging ; etiology ; Low Back Pain ; etiology ; Lumbar Vertebrae ; Male ; Nerve Fibers ; pathology ; Rabbits ; Radiography ; Random Allocation ; Ubiquitin Thiolesterase ; metabolism

OBJECTIVETo analyze the correlation between the genotype and phenotype of 18q deletion syndrome with chromosome microarray analysis (CMA).

METHODSEight cases with 18q deletion syndrome were selected, including two affected fetuses and six children patients. DNA was extracted and hybridized with Affymetrix CytoScan TM 750K arrays following the manufacturer's standard protocol. The data was analyzed with a special software package.

RESULTSCMA analysis identified pathogenic copy number variations (CNVs) on 18q in all cases, which ranged from 6.612 Mb to 22.973 Mb. NFATC1, GALR1, MBP, SALL3 and TSHZ1 are likely to be causative genes for congenital heart disease, psychological, growth retardation, and cleft palate.

CONCLUSIONCMA can precisely locate the breakpoints of 18q and facilitate definition of the genotype-phenotype correlations, which is useful for prognosis.

Child, Preschool ; Chromosome Deletion ; Chromosome Disorders ; genetics ; Chromosomes, Human, Pair 18 ; genetics ; DNA Copy Number Variations ; Female ; Humans ; Infant ; Male ; Microarray Analysis

Background/Aims: Esophagogastric junction adenocarci-noma (EJA) is a malignant tumor associated with high mor-bidity and has attracted increasing attention due to a rising incidence and low survival rate. Pathological biopsy is the gold standard for diagnosis, but noninvasive and effective tests are lacking, resulting in diagnoses at advanced stages.This study explored the diagnostic value of insulin-like growth factor binding protein 7 (IGFBP7) in EJA. Methods: A total of 120 EJA patients and 88 normal controls were recruited, and their serum levels of IGFBP7 were measured by enzymelinked immunosorbent assay. Receiver operating character-istic (ROC) curve analysis was used to assess the diagnostic value, and Pearson chi-square analysis was used to evaluate the correlation between IGFBP7 and clinical parameters. Ka-plan-Meier survival analysis was carried out to assess the ef-fect of IGFBP7 on overall survival (OS). Results: The levels of IGFBP7 were higher in both early- and late-stage EJA patients than in normal controls (p<0.001). The area under the ROC curve for EJA patients was 0.794 (95% confidence interval [CI], 0.733 to 0.854), with a cutoff value of 2.716 ng/mL, a sensitivity of 63.3% (95% CI, 54.0% to 71.8%) and a specific-ity of 90.9% (95% CI, 82.4% to 95.7%). For the diagnosis of early-stage EJA, the same cutoff value and specificity were obtained, but the sensitivity of IGFBP7 was 54.3% (95% CI, 36.9% to 70.8%). Patients with low IGFBP7 protein expres-sion had lower OS than those with high expression (p=0.034).The multivariate analysis showed that IGFBP7 is an inde-pendent prognostic factor for EJA (p=0.011). Conclusions Serum IGFBP7 acts as a potential diagnostic and prognostic marker for EJA.

A new method for simultaneous determination of 23 kinds of per-and polyfluoroalkyl substances(PFASs)(13 kinds of perfluoro carboxylic acids,4 kinds of perfluoro sulfonic acids,and 6 kinds of new substitutes)in plant leaf tissue by ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)using automatic online solid phase extraction(SPE)to remove the matrix interference components in plant crude extracts was developed.The plant leaf samples were extracted twice with 1%formic acid-methanol solution,then evaporated to dry,redissolved with 70%methanol solution,and directly injected for analysis.After 23 kinds of target PFASs were purified automatically by online SPE with a WAX column,the six-way valve was switched to rinse PFASs onto an alkaline mobile phase system-compatible C18 analytical column.Then,the 23 kinds of target PFASs were separated within 16 min by gradient elution using a binary mobile phase system of methanol/water(Containing 0.4%ammonium hydroxide).Tandem mass spectrometry was performed in multiple reaction monitoring(MRM)mode for online detection of various PFASs,and quantification was carried out by internal standard method.The results of the method validation showed that satisfactory average recoveries of 23 kinds of PFASs in plant leaf samples(64.2%-125.5%),precision(relative standard deviations(RSDs)of 0.7%-12.8%),linearity(R2＞0.990),and sensitivity(the detection limits(S/N=3)were in the range of 0.02-0.50 μg/kg)were achieved.Finally,this method was used to detect PFASs in the marine green tide algae(Enteromorpha prolifera)and several tree leaves,and a total of 6 kinds of PFASs were detected,in which PFBA was the main contaminant.Compared with the reported offline SPE methods,the proposed online SPE technique significantly simplified the sample pretreatment process and provided an automatic,simple,and environment-friendly method for the routine monitoring of legacy and emerging PFASs in plant tissues.