Objective To investigate the correlation between neonatal infectious disease and brain injury.MethodsClinical data of 1266 newborns with infectious diseases were collected from Peking University First Hospital from November 2005 to August 2010.The occurrence of brain injury was summarized.Related factors of brain injury caused by infection and the risk factors for severe brain injury were analyzed by Logistic regression model. Results Among the newborns with neonatal infectious diseases, the incidence of brain injury was 8.6％(108/1266), including 101 (8.0％)mild cases and seven (0.6％) severe cases. The incidence of brain injury for the newborns with severe infectious diseases was higher than those with mild infectious diseases [38.7％(29/75) vs 6.7％(79/1191),x2=92.787,P=0.000].The incidence of brain injury for the newborns withobviousinflammatoryreactionwassignificantlyhigherthanthosewithout [(13.0％(26/200) vs 7.5％ (77/1025),x2=6.544,P=0.011].Severe infection was independent risk factor for severe brain injury by Logistic regression model analysis (OR =15.750,95％ CI:1.756-141.281,P=0.014).ConclusionsIniectious diseases could cause injury on central nervous system,especially when there are severe infections or inflammatory reactions. The severer the infection,the severer the brain injury,especially when complicated by some factors such as asphyxia and hypoglycemia.

Objective To investigate the value of early quantified analysis of perinatal white matter injury by cranial ultrasound gray scale measurement. MethodsThe cranial ultrasound exam was performed in 152 newborns with different gestational age0 early after their birth. These newborns were divided into two groups: 104 newborns diagnosed as white matter injury within 7 days after birth were taken as patient group; while 48 newborns who were not were taken as control group. The gray scale values in the trigone of lateral ventricle of white matter were analyzed by medical image analysis system. The newborns in patient group accepted cranial ultrasound exam at one month after birth, the grey scale value and cyst in the white matter were recorded. Three to six months old, the cranial ultrasound exam was repeated to record the change of white matter volume, morphology of lateral ventricle and change of the cysts. When they were 1.5 to 2 years old, the neurological function were quantitatively evaluated with Gesell score, and the results were classified as normal and abnormal.The relationships between gray scale value and neuro-developmental outcome were analyzed with receiver operating characteristic curve.Results During neonatal period, the average gray scale values in severely injured group was 131.72±2.40, higher than that of mildly injured group (116.61±2.48), and which in mildly injury group was higher than that in control group (100.50±1.66) (q=4. 521 and 4. 492, P＜0. 05). It was showed by receiver operating characteristic curve that gray scale value ＞114.37 could help to diagnose white matter injury, with the sensitivity of 0. 721 and the specificity of 0. 854; gray scale value ＞119.80 could help to diagnose severe white matter injury,with the sensitivity of 0. 716 and the specificity of 0. 776.As the gray scale value increased, the incidence of white matter volume decreased and the enlargement of lateral ventricle in the later period of injury increased. Patients with gray scale value ＞ 130 tended to suffer from leucomalacia. During neonatal period, the incidence of abnormal neurodevelopment before 2 years old was 5.0％ in patients with gray scale value ＜ 110, while it was 27.8 ％ in the patients with gray scale value between 110 and 120, 47.8％ in the patients with gray scale value ＞ 120.Conclusions Quantified analysis of ultrasound gray scale value might be promising in early diagnosis of perinatal white matter injury through early judgement of the outcomes of white matter injury and forward neurodevelopment.

Postoperative hydrocephalus is one of the most common complications in resection of lateral ventricle trigone tumor, which can seriously affect the prognoses of patients. Possible influencing factors include tumor resection degrees, postoperative meningitis, intraoperative intraventricular hemorrhage, and cerebrospinal fluid drainage tube placement, but the specific mechanism is still unclear. In this paper, the influencing factors and possible mechanisms of hydrocephalus after lateral ventricle trigone tumor are reviewed as follows.

Objective To investigate the value of clinical follow-up in prenatal diagnosis of isolated double aortic arch (DAA) . Methods The clinical follow-up materials were retrospectively reviewed in 17 fetuses . Of all the isolated DAA fetuses ,the accuracy rate of prenatal diagnosis was confirmed by CT ,MRI , autopsy or echocardiography ,and pregnant outcomes were summarized . Results A total of 17 fetuses had a sonographic diagnosis of isolated DAA in our centers at a mean gestational age of 23 -32(27 ± 3) weeks , with mother mean age 19 -44 (28 ± 6) years old . One case of DAA type-A was misdiagnosed ,15 cases were delivered with 2 cases occurred respiratory distress or mild dysphagia ,who received surgical treatment , and 13 cases clinical findings were unremarkable at the fellow-up of 24 months ,the silent-rate of clinical symptoms was 86％ . Termination of pregnancy happened in 2 cases ,with 1 (6％ ) had additional ventricular septal defects ,another ( 6％ ) had additional anomalies of congenital high airway obstruction syndrome . In 17 cases of fetuses with isolated DAA ,there were 15 ( 88％ ) cases with dominant right-sided arch ,1 case (6％ ) with dominant left arch ,and 1 case (6％ ) with equal arches in size .Karyotyping prenatal testing was offered to 15 parents with normal results . Conclusions Prenatal ultrasound can accurately diagnose isolated DAA by multiple sections scan ,expanding diagnostic ideas by combination with other medical imaging data to prevent apparent life-threatening event ,or to guide for ex-utero intrapartum treatment . Isolated DAA clinical follow-up results in good outcome .

OBJECTIVEThe aim of this study was to establish a standard protocol for detection of EGFR mutations in cytologic specimens.

METHODS287 cytologic samples were collected from the patients who were suspected of having lung cancer at six hospitals in Beijing. A detection protocol for EGFR mutations was designed. Two comparative experiments were carried out for the coincidence in EGFR mutation rates between direct sequencing (Seq) and amplification refractory mutation system (ARMS) methods, and between 40 matched cytologic samples with formaldehyde-fixed paraffin embedded (FFPE) cytologic blocks and cytospin slides.

RESULTSTumor cells were found in 236 out of 287 cases (82.2%, 236/287) . Among them, there were 31 cases (13.1%, 31/236) of low tumor cell content samples and 205 cases (86.9%, 205/236) of high tumor cell content samples. 180 cases in the high tumor cell content samples (87.8%, 180/205) were diagnosed to be consistent with NSCLC. 25 out of 194 cases were ruled out or indefinite to be diagnosed as NSCLC by immunohistochemistry. By direct sequencing, the mutation rate of EGFR was 27.8% (50/180) in NSCLC samples and 28.2% (50/177) in adenocarcinoma samples (high tumor content samples) . By ARMS, the mutation rate of EGFR was 45.6% (82/180) in NSCLC samples and 46.3% (82/177) in adenocarcinoma samples (high tumor content samples). The EGFR mutation rate in low tumor content samples was 38.7% (12/31) , there was no significant difference in EGFR mutation rates between the groups of low tumor cell content samples and high tumor cell content samples (P = 0.12). The concordance rate of EGFR mutation rates was 100% between scraping tumor cells from slides samples and from FFEP blocks in the 40 matched samples. Forty-eight out of 180 definitive NSCLC patients received Gefitinib therapy. The FPS was 12 months in the gefitinib-treated ARMS⁺ group and 2 months in the ARMS⁻ group (P < 0.001), and the OS was 19 months in the gefitinib-treated ARMS⁺ group and 7 months in the ARMS⁻ group (P = 0.003), but no significant differences were found in the efficacy (PFS and OS) of Gefitinib between Seq⁺ and Seq⁻ groups (P = 0.227, P = 0.510, respectively), and Seq⁺/ARMS⁺ and Seq⁻/ARMS⁺ groups (P = 0.354, P = 0.334, respectively).

CONCLUSIONSThe detection protocol for EGFR mutations in cytological specimens introduced in this study is tested to be reliable and feasible. Pathological evaluation and immunohistochemistry are important in the detection procedure of EGFR mutations in cytologic specimens. High sensitivity methods should be selected for detection of EGFR mutations in cytologic samples.

Adenocarcinoma ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Humans ; Lung Neoplasms ; diagnosis ; epidemiology ; metabolism ; Mutation ; Mutation Rate ; Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; metabolism

Objective To explore the value of prenatal ultrasound in diagnosis of omphalocele-exstrophy-imperforate anusspinal defects (OEIS) in first trimester.Methods Prenatal ultrasonic characteristics of 10 fetuses with OEIS complex in first trimester were retrospectively analyzed and compared with autopsy results.Results Cystic bulging in the lower anterior abdominal wall was observed in all 10 fetuses.Spinal scoliosis dysplasia was found in 10 fetuses,with myelomeningocele in 3 fetuses.No normal bladder was visualized in 8 fetuses.Thickened nuchal translucency was noticed in 5 fetuses,among which neck lymphatic hydrocele was found in 1 fetus.The bilateral clubbed feet and left lower mutilation was observed in 1 fetus,respectively.All 10 OEIS complex fetuses were found accompanied with short umbilical cord,while single umbilical artery and umbilical cord cyst were found in 4 and 1 fetus,respectively.Autopsy showed abdominal wall defects with exstrophy in 10 fetuses.However,no complete cystic bulging was found.Besides,autopsy also showed pubic symphysis separation and bladder exstrophy in 10 fetuses without obvious genitalia nor anus.Conclusion Cystic bulging in the lower anterior abdominal wall is the most common prenatal ultrasonic characteristic of OEIS complex in first trimester.

OBJECTIVEThe aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens.

METHODSLung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical (IHC) ALK(D5F3) detecting ALK protein expression was performed in 203 prepared formalin-fixed paraffin-embedded (FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid (BL) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization (FISH). Six patients with ALK IHC-positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments: (1) Comparison of the results of 4% neutral buffered formalin fixed for different time (24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks; (2) Comparing qRT-PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens.

RESULTSAmong the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results (by at least one method), with an ALK test ratio of 90.4% (207/229). FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK (D5F3) were successfully performed in all the 203 FFPE cell blocks (100%), and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT-PCR, and 96 out of 98 (97.96%) cytologic samples were successfully performed.18 out of 19 IHC ALK-positive cases were verified to be of ALK fusion status by qRT-PCR. The concordance rate was 94.7% (Kappa=0.967, P<0.001) between Ventana IHC ALK (D5F3) and qRT-PCR, and the sensitivity of the Ventana IHC ALK (D5F3) assay compared with qRT-PCR was 100% and the specificity was 98.7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK (D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK (D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT-PCR test and ALK gene fusion showed good concordance.

CONCLUSIONSThe standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK (D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK (D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT-PCR may be an alternative option for the detection of ALK gene fusion.

Adenocarcinoma ; drug therapy ; enzymology ; genetics ; pathology ; Alkaline Phosphatase ; genetics ; metabolism ; Gene Fusion ; Genes, erbB-1 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; drug therapy ; enzymology ; genetics ; pathology ; Protein Kinase Inhibitors ; therapeutic use ; Proteomics ; Pyrazoles ; therapeutic use ; Pyridines ; therapeutic use ; Sensitivity and Specificity

Objective: To establish a triple-color pseudovirion-based neutralization assay (PBNA) and evaluate its capability of detecting immunogenicity of the sera generated by the immunization of HPV 9-valent vaccine. Methods: HPV pseudovirus (PsVs) 6/11/16/18/31/33/45/52/58 with the encapsidated fluorescence expressing red fluorescent plasmid N31-MCHREEY, green fluorescent N31-EGFP or blue fluorescent N31-mTagBFP were generated. The concentration of HPV PsVs and the infection titers of HPV PsVs were detected by double-antibody sandwich ELISA and TCID₅₀, respectively. The single- and triple color HPV 16/33/45 PsVs were used to detect the neutralization titers of mice sera immunized with HPV 9-valent vaccine and confirmed the accuracy and specificity of the triple-color PBNAs. Then, the single- and triple color HPV 6/11/18/31/33/45/52/58 PsVs were employed to detect the neutralization titers of cynomolgus macaques sera immunized with HPV 9-valent vaccine and determined whether the triple-color PBNAs could be applied to evaluate the immunogenicity of the sera generated by the immunization of HPV9-valent vaccine. Results: The concentration of HPV16 PsVs encapsulating green, red or blue fluorescent plasmid was 5.0 to 6.0 μg/ml and HPV6/11/18/31/33/45/52/59 triple-color HPV PsVs was about 1.0 to 3.0 μg/ml. 9 types HPV PsVs containing EGFP, Mcherry or mTagBFP reporter plasmid were obtained and the concentration can meet the need of neutralization detection. 9 types single-color fluorescent HPV PsVs had similar infectivity against 293FT cells with the infection titer values between 1×10⁴ and 1×10⁵. The results of PBNAs showed that there was no significant difference in the anti-HPV neutralization titers of mice sera induced by HPV 9-valent vaccine between single-color and triple-color HPV16/33/45 PsVs (P>0.05). Similarly, there was also no significant difference in the anti-HPV neutralization titers of cynomolgus macaques sera induced by HPV 9-valent vaccine between single-color and triple-color HPV6/11/18/31/33/45/52/58 PsVs (P>0.05). Conclusion We successfully established the triple-color PBNAs and verified the accuracy and specificity of triple-color PBNAs consistent with single-color PBNAs. The triple-color PBNAs can be applied to evaluate the immunogenicity of HPV 9-valent vaccine's immune serum.

OBJECTIVES: The aim of this study was to compare the anterior and posterior occlusal plane characteristics of patients with different temporomandibular joint osseous statuses. METHODS: A total of 306 patients with initial cone beam CT (CBCT) and cephalograms were included. They were divided into three groups on the basis of their temporomandibular joint osseous status: bilateral normal (BN) group, indeterminate for osteoarthrosis (I) group, and osteoarthrosis (OA) group. The anterior and posterior occlusal planes (AOP and POP) of the different groups were compared. Then, the regression equation was established after adjusting for confounding factors, and a correlation analysis between the occlusion planes and other parameters was performed. RESULTS: SNA, SNB, FMA, SN-MP, Ar-Go, and S-Go were correlated with the occlusal planes. Relative to the BN and I groups, the FH-OP of the OA group increased by 1.67° on the average, FH-POP increased by 1.42° on the average, and FH-AOP increased by 2.05° on the average. CONCLUSIONS The occlusal planes were steeper in the patients with temporomandibular osteoarthrosis than in the patients without it, and the mandible rotated downward and backward. The height of the mandibular ramus, the mandibular body length, and the posterior face height were small. In clinical practice, attention should be given to the potential risk of temporomandibular joint osteoarthrosis in such patients. In addition, SNB, FMA, SN-MP, Ar-Go, S-Go, and occlusal planes had moderate correlations.
Humans ; Dental Occlusion ; Cephalometry ; Mandible ; Temporomandibular Joint Disorders/diagnostic imaging* ; Temporomandibular Joint/diagnostic imaging* ; Osteoarthritis/diagnostic imaging* ; Mandibular Condyle