Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; Humans ; Infant ; Infant, Newborn ; Male ; Young Adult

Objective The purpose of this study was to develop a high-throughput micro-plate radiobinding assay (RBA) of glutamic acid decarboxylase antibody (GAD-Ab) and to evaluate its clinical application. Methods 35labeled GAD65 antigen was incubated with sera for 24 h on a 96-well plate, and then transferred to the Millipore plate coated with protein A, which was washed with 4℃ PBS buffer, and then counted by a liquid scintillation counter. The GAD-Ab results were expressed by WHO standard unit (U/ml). A total of 224 healthy controls, 162 patients with type 1 diabetes mellitus(T1DM) and 210 patients with newly diagnosed type 2 diabetes (T2DM) were recruited. A total of 119 TI DM and healthy cases with gradually changing GAD-Ab levels were selected to compare the consistency of micro-plate RBA with conventional radioligand assay (RLA). Blood samples were obtained from the peripheral vein and finger tip in 32 healthy controls, 35 T1DM and 24 T2DM patients, and tested with micro-plate RBA and then compared with the conventional RLA to investigate the reliability of finger tip sampling. Linear correlation,student's t-test, variance analysis and receiver operating characteristic (ROC) curve were performed using SPSS 11.5. Results (1) The optimized conditions of micro-plate RBA included 2 μl serum incubated with3 ×104 counts/min 35S-GAD for 24 h under slow vibration, antigen-antibody compounds washed 10 times by 4℃ PBS buffer, and radioactivity counted with Optiphase Supermix scintillation liquid. (2)The intra-batch CV of the micro-plate RBA was 3.8%- 10.2%, and the inter-batch CV was 5.6%- 11.9%. The linearity analysis showed a good correlation when the GAD-Ab in serum samples ranged from 40.3 to 664 U/ml and the detection limit of measurement was 3.6 U/ml. The results from Diabetes Autoantibody Standardization Program (DASP) 2005 showed that the sensitivity and specificity for GAD-Ab were 78% (39 positive among 50 new-onset T1DM) and 98% (2 positive among 100 healthy controls). The results of GAD-Ab obtained with micro-plate RBA and RLA were closely correlated (r=0.915,P<0.001) with a high concordance level of 97.5% and a Kappa value of 0.95. (3)TI DM and T2DM patients showed higher positive rates for GAD-Ab than the healthy controls(46.9% and 5.2% vs 0.89% ,X2=123.5 and 10. 1 ,P <0.001 and <0.01, respectively). (4)The consistency of GAD-Ab measurement with RBA using finger tip blood and RLA measurement using venous blood was 96.7% (r =0.946,P <0.001, Kappa value: 0.905). Conclusions The micro-plate RBA of GAD-Ab has high sensitivity, specificity and reproducibility, and can be measured with finger tip blood sampling. It might be a better alternative for clinical practice.

OBJECTIVE: To observe the expression of aquaporin 4 (AQP4) in diffuse brain injury (DBI) of rats and to explore the corresponding effect of AQP4 for brain edema. METHODS: The rat model of DBI was established using Marmarou's impact-compression trauma model. Brain water content was measured by dry-wet weight method. Blood-brain barrier permeability was evaluated by Evans blue (EB) staining. Immunohistochemical method was used to observe the expression of AQP4. RESULTS: Brain water content increased after 3 h and peaked at 24 h after DBI. Brain EB content significantly increased and peaked at 12 h after DBI. The expression of AQP4 significantly increased after 3 h and peaked at 24 h after DBI, and the number of AQP4 positive astrocytes increased. CONCLUSION The increment of the permeability of blood-brain barrier and the expression of AQP4 may contribute to the development of brain edema in rat DBI. The change of AQP4 expression in astrocytes may also contribute to determine DBI.
Animals ; Aquaporin 4/metabolism* ; Astrocytes ; Blood-Brain Barrier/metabolism* ; Brain ; Brain Edema/metabolism* ; Brain Injuries/metabolism* ; Cell Membrane Permeability/genetics* ; Disease Models, Animal ; Permeability ; Rats ; Water

OBJECTIVE: To investigate the risk factors for hypoglycemia after birth in preterm infants with a gestational age of ≤32 weeks. METHODS: A retrospective analysis was performed for 86 neonates with hypoglycemia and a gestational age of ≤32 weeks who were admitted to the neonatal intensive care unit from January 2017 to June 2020 (hypoglycemia group). A total of 172 preterm infants with normal blood glucose who were hospitalized during the same period were randomly enrolled as the control group. Univariate analysis and multivariate logistic regression analysis were used to screen out the risk factors for hypoglycemia in preterm infants. RESULTS: There were 515 preterm infants during the study, among whom 86 (16.7%) had hypoglycemia. Compared with the control group, the hypoglycemia group had significantly higher percentages of small for gestational age (SGA), cesarean section, maternal hypertension, and antenatal steroid administration (P<0.05), but significantly lower birth weight and rate of intravenous glucose use before blood glucose test (P<0.05). SGA (OR=4.311, 95%CI: 1.285-14.462, P<0.05), maternal hypertension (OR=2.469, 95%CI: 1.310-4.652, P<0.05), and antenatal steroid administration (OR=6.337, 95%CI: 1.430-28.095, P<0.05) were risk factors for hypoglycemia in preterm infants, while intravenous glucose use (OR=0.318, 95%CI: 0.171-0.591, P<0.05) was a protective factor against hypoglycemia in preterm infants. CONCLUSIONS SGA, maternal hypertension, and antenatal steroid administration may increase the risk of early hypoglycemia in preterm infants with a gestational age of ≤32 weeks, and intravenous glucose use is recommended as soon as possible after birth for preterm infants with a gestational age of ≤32 weeks to reduce the incidence rate of hypoglycemia.
Cesarean Section ; Female ; Gestational Age ; Humans ; Hypoglycemia/etiology* ; Infant ; Infant, Newborn ; Infant, Premature ; Infant, Small for Gestational Age ; Pregnancy ; Retrospective Studies ; Risk Factors

Objective To evaluate the association of islet autoantibodies [ glutamic acid decarboxylase antibody(GADA),protein tyrosine phosphatase antibody(IA-2A)and insulin autoantibodies(IAA)1 with HLA- DQ genotypes in the first-degree relatives of autoimmune type 1 diabetes mellitus.Methods This was a cross- sectional and case-control study.Three hundred and fifty-one first-degree relatives with normal glucose tolerance of patients with type 1 diabetes mellitus and 376 healthy controls were recruited and measured for GADA,IA-2A and IAA by radioligand assay,and 156 first-degree relatives of patients with autoimmune type 1 diabetes mellitus and 278 controls were typed for genetic polymorphisms of HLA-DQ with PCR sequencing-based typing method.Results (1)DQA1*03,DQBI*0303,*0401 alleles and DQA1 * 03-DQBI * 0303,DQA1 * 05-DQBI * 0201,DQA1 * 03-DQBI * 0401 haplotypes were significantly increased in the first-degree relatives of autoimmune type 1 diabetes mellitus(P

OBJECTIVETo explore CT and MRI manifestations of the axial area peripheral primitive neuroectodermal tumors (pPNETs) in order to improve the knowledge of this disease.

METHODSThe clinical data of 10 patients with pPNETs underwent pathologically confirmed were retrospectively analyzed from October 2008 to May 2014. There were 7 males and 3 females, aged from 8 to 49 years old with median of 23.6 years. The preoperative multi-slice spiral CT scan was completed in 3 cases, plain CT scan and enhancement in 4 cases; MRI and enhancement scanning in 5 cases; and among them, 2 cases underwent both MRI and CT scan.

RESULTSIn-bone type was found 6 cases and out-bone type was found 4 cases. Three cases occurred in sacral vertebrae, 2 cases in lumbar vertebrae, 1 case in cervical vertebrae, 1 case in cervical spinal canal, 1 case in coccyx, 1 case in the right iliac bone, 1 case in presacral space. Cross sectional the smallest tumor maximum level was 1.1 cmx 1.2 cm in size, the biggest tumor was 8.0 cm x 9.2 cm, the median size was 4.4 cm x 5.7 cm, of them, the tumor of maximal diameter larger than 5 cm had 6 cases. Except 2 cases-without destruction of bone, the other 5 cases with osteolytic destruction, 2 cases with calcification, 1 case with mixed. Equidensite was main in CT scan, 1 case with uniform density, other 6 cases with uneven density,in which 3 cases with "floating ice" change; 1 case with moderate strengthening, other 3 cases with obviously strengthening, 2 cases with multiple small blood vessels in enhancement scanning. MRI of 5 cases showed the signal of isointensity on T1WI, the slightly high signal on T2WI and the signal was not uniform; after enhancement scan, the signal of 5 cases obviously enhanced. Two patients complicated with vertebral compression fractures, no periosteal reaction was found in all patients, and no the destruction of intervertebral disk was found in 5 patients of MRI scan.

CONCLUSIONThe axial area pPNETs is common among children and the youth, and the mass often is huge. The mass of in-bone type often envelopes the vertebral body, and main located on prevertebral space, all associated with bone destruction, osteolytic destruction is common, and primary vertebral bodies also is common, attachment primary or involvement is few found, it can involve the spinal canal and anterior wall of spinal canal is common, some cases complicate with multiple newly born small vessels. The mass of out-hone type in deep soft tissue is common, minority primary spinal canal, many complicated with vertebral bone destruction, osteolytic destruction was main. The intervertebral disk was not invaded and intervertebral space has not stenosis. CT scan offer complicate with "floating ice" sign, and in-bone type is common. Isointensity is main on MRI TlWI and slightly longer signal is main on MRI T2WI, strengthening signal is obvious.

Adolescent ; Adult ; Child ; Cross-Sectional Studies ; Diagnosis, Differential ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neuroectodermal Tumors, Primitive, Peripheral ; diagnosis ; diagnostic imaging ; Radionuclide Imaging ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo investigate the feasibility of construction of tissue-engineered skin in vitro.

METHODSFibroblasts were seeded on the dermal surface of acellular dermal matrix (ADM). 7 days later, epidermal cells (5 x 10(5)/cm2) were mixed with fibroblasts (0.2 x 10(5)/cm2) and then seeded on epidermal surface of ADM. The culture medium was the mixture liquor containing K-SFM in half and the culture supernatants of fibroblasts in half. In the control group, only epidermal cells (5 x 10(5)/cm2) were seeded and cultured with K-SFM. After composite skin was cultured for 1 week and 3 week, samples were harvested respectively for morphological study and to receive identification by immunohistochemistry.

RESULTSAfter 3-week culture, there were 3-4 continuous layers of cells in the epidermis in the experiment group. The epidermis was attached tightly to the dermis with trochanterellus. But in the control group, there were just 1-2 layers of cells in the epidermis which was not connected to the dermis. Strong positive dye of Laminin indicated that basement membrane was thoroughly formed in the experiment group. The same result was demonstrated by transmission electron microscope.

CONCLUSIONSThe mix-seeding of epidermal cells and few fibroblasts would promote the adhesion and proliferation of epidermal cells on ADM, and the formation of basement membrane.

Animals ; Cells, Cultured ; Coculture Techniques ; Epidermis ; cytology ; Fibroblasts ; cytology ; Skin, Artificial ; Swine ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo investigate the preemptive analgesic efficiency of parecoxib in patients undergoing laparoscopic colorectal surgery.

METHODSForty ASA I-II patients aged 30 to 64 years undergoing laparoscopic colorectal surgery were randomized to receive either intravenous parecoxib sodium (40 mg) at anesthesia induction (group A) or intravenous parecoxib sodium (40 mg) 30 min before the completion of surgery (group B). Butorphanol was administered by patient-controlled analgesia for postoperative analgesia. The intensity of pain measured by VAS score was recorded at 2, 4, 6, 8, 12, and 24 h after the operation. The number of unsatisfied demand and the number of successfully delivered doses, butorphanol consumption at 12 h and 24 h after the operation, the patients' global evaluation of the postoperative analgesia and the number of the patients receiving rescue medication and adverse effects related to analgesia were recorded and compared between the two groups.

RESULTSThe VAS scores at different time points were significantly lower in group A than in group B (P<0.05). The number of unsatisfied demand and the number of successfully delivered doses were significantly higher and butorphanol consumption at 12 h and 24 h after the operation was significantly less in group A (P<0.05). The incidence of adverse events was similar between the two groups.

CONCLUSIONAdministration of 40 mg parecoxib sodium at anesthesia induction in the patients undergoing laparoscopic colorectal surgery can result in significant preemptive analgesia.

Adult ; Analgesia ; methods ; Analgesia, Patient-Controlled ; Analgesics, Opioid ; administration & dosage ; Butorphanol ; administration & dosage ; Colorectal Surgery ; Humans ; Isoxazoles ; administration & dosage ; Laparoscopy ; Middle Aged

OBJECTIVETo study the efficacy of epidural block combined with general anesthesia on stress reaction in spontaneous hypertensive rats (SHR).

METHODSEighteen SHR and 18 SD rats (above 24 weeks) were randomly divided into 6 groups, namely epidural block combined with general anesthesia group (A1,A2), general anesthesia group (B1,B2) and control group (D1,D2). General anesthesia was performed with celiac injection of droperidol, fentanyl and diazepam. Tracheal intubation and ventilation were performed after tracheotomy, and epidural block was conducted by incision. The rats in groups A1, A2, B1, B2 underwent splenectomy. All the rats were sacrificed 24 h after the surgery and two myocardium specimens were collected for detecting c-fos and HSP70 expression using RT-PCR.

RESULTSc-fos mRNA expression was significantly lower but HSP70 mRNA expression significantly higher in group A1 than in group B1.

CONCLUSIONCompared with general anesthesia, general anesthesia combined with epidural block can reduce the stress responses and protect cardiac myocytes by decreasing c-fos expression and increasing HSP70 expression in SHR.

Anesthesia, Epidural ; Anesthesia, General ; Animals ; HSP70 Heat-Shock Proteins ; metabolism ; Myocardium ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Rats, Inbred SHR

OBJECTIVETo study the diagnostic value of diffusion tensor imaging (DTI) in cervical spondylotic myelopathy.

METHODSTwenty healthy volunteers and fifty patients with cervical spondylotic myelopathy underwent DTI in the Affiliated Hospital of Medical College of Ningbo University from January 2014 to April 2015. Healthy volunteers served as controls. Fifty patients were divided into three groups (group A , B, C) according to cervical MRI scan standard. Group A (17 cases) had only the dura mater spinalis compressed; Group B (23 cases) showed the cervical spinal cord compressed, but no high signal in it; Group C (10 cases) had the cervical spinal cord compressed with high signal in the same level. The average apparent diffusion coefficients(ADC) and fractional anisotropy (FA)values in these examinee were analyzed and all subjects were performed fiber tracking.

RESULTSThere was no statistically significant differences in ADC and FA values in C2/C3, C3/C4, C4/C5, C5/C6, C6/C7 of control group (P>0.05). The average ADC and FA values in control group were (0.875 +/- 0.096) x10(3) mm2/s and 0.720 +/- 0.051, respectively; compared with group A,there was no statistically significant difference; compared with group B and C, there was significant difference; comparison among group A, B, C, there was significant differences.

CONCLUSIONDTI can early and accurately quantify the changes of microstructure in cervical spondylotic myelopathy. Fiber tracking can show the damage range of spinal cord lesions.

Adult ; Case-Control Studies ; Cervical Vertebrae ; diagnostic imaging ; Diffusion Tensor Imaging ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Spinal Cord Diseases ; diagnostic imaging ; surgery ; Spondylosis ; diagnostic imaging ; Young Adult