Clinical Value of Hybrid TOF-PET/MR Imaging-Based Multiparametric Imaging in Localizing Seizure Focus in Patients with MRI-Negative Temporal Lobe Epilepsy.

BACKGROUND AND PURPOSE: Temporal lobe epilepsy is the most common type of epilepsy. Early surgical treatment is superior to prolonged medical therapy in refractory temporal lobe epilepsy. Successful surgical operations depend on the correct localization of the epileptogenic zone. This study aimed to evaluate the clinical value of hybrid TOF-PET/MR imaging-based multiparametric imaging in localizing the epileptogenic zone in patients with MR imaging-negative for temporal lobe epilepsy. MATERIALS AND METHODS: Twenty patients with MR imaging-negative temporal lobe epilepsy who underwent preoperative evaluation and 10 healthy controls were scanned using PET/MR imaging with simultaneous acquisition of PET and arterial spin-labeling. On the basis of the standardized uptake value and cerebral blood flow, receiver operating characteristic analysis and a logistic regression model were used to evaluate the predictive value for the localization. Statistical analyses were performed using statistical parametric mapping. The values of the standardized uptake value and cerebral blood flow, as well as the asymmetries of metabolism and perfusion, were compared between the 2 groups. Histopathologic findings were used as the criterion standard. RESULTS: Complete concordance was noted in lateralization and localization among the PET, arterial spin-labeling, and histopathologic findings in 12/20 patients based on visual assessment. Concordance with histopathologic findings was also obtained for the remaining 8 patients based on the complementary PET and arterial spin-labeling information. Receiver operating characteristic analysis showed that the sensitivity and specificity of PET, arterial spin-labeling, and combined PET and arterial spin-labeling were 100% and 81.8%, 83.3% and 54.5%, and 100% and 90.9%, respectively. When we compared the metabolic abnormalities in patients with those in healthy controls, hypometabolism was detected in the middle temporal gyrus (P < .001). Metabolism and perfusion asymmetries were also located in the temporal lobe (P < .001). CONCLUSIONS: PET/MR imaging-based multiparametric imaging involving arterial spin-labeling may increase the clinical value of localizing the epileptogenic zone by providing concordant and complementary information in patients with MR imaging-negative temporal lobe epilepsy.

[Clinical Application Value of Peripheral Blood Diagnostic Report].

Objective: To explore the clinical application value of peripheral blood diagnostic report. Methods: 557 peripheral blood diagnostic reports were collected from Peking University First Hospital, YANDA LU DAOPEI Hospital and Beijing United Family Hospital. The results were analyzed and summarized according to different blood cell morphology character for the first time and review cases, respectively. Results: Two hundred and one samples from first time patients were found abnormal complete blood count or leukocyte differential count, they were summarized as anemia, anemia accompanied with leukopenia or thrombopenia, abnormal white blood cell count or leukocyte differential count and abnormal platelet count. Each condition was further distinguished on the basis of different morphology character. Initial diagnosis or further examination could be proposed if abnormal morphology was specific or typical, when blood cell morphology was atypical or normal, the morphology was described objectively. 22 review cases included many benign and malignant disorders such as acute leukemia, chronic leukemia, myelodysplastic syndrome, multiple myeloma, infectious mononucleosis and so on. Suggestion of therapeutic effect, progression of diseases or further examination could be present according to complete blood cell count and morphology character. Conclusion: Peripheral blood diagnostic report can provide more comprehensive and accurate information for clinic, and propose important advisory opinions for primary diagnosis, differential diagnosis, treatment monitoring and progression assessment.

[Clinical effects of Dynesys system and transfacet decompression through Wiltse approach in the treatment of lumbar degenerative diseases].

Objective: To investigate the early clinical effects of Dynesys system and transfacet decompression by Wiltse approach in the treatment of lumbar degenerative diseases. Methods: From January 2010 to December 2013, 48 patients suffering from lumbar degenerative diseases were treated with Dynesys system in addition to transfacet decompression through Wiltse approach.There were 28 males and 20 females with age of (51.8±6.8). The preoperative diagnosis included lumbar spinal stenosis(10 cases); lumber intervertebral disc herniation (38 cases). There were 23 cases in L4/5, 16 cases in L5/S1 and 9 cases in both of L4/5 and L5/S1.Posterolateral fixation with Dynesys pedicle screw through Wiltse approach.Unilateral resection of the inferior articular facet of the superior vertebra and the superior articular facet of the inferior vertebra.Decompression of the vertebral canal until the never root was decompressed satisfactorily.In the end, Dynesys was performed according to normal procedure.VAS, ODI evaluating standards were applied to evaluate the therapeutic effect.The intervertebral space and ROM of the lumbar were observed by X ray. Results: All patients underwent surgery safely without severe complications occurred.The average following up time was 33.5 (24-60) months.Compared with preoperative parameters (7.7±1.3, 70.8±13.5), the scores of VAS and ODI decreased significantly after surgery (2.3±1.5, 23.6±12.2) and at the final follow-up (2.2±1.4, 20.0±9.8) (P<0.05). There were significant difference in the height of intervertebral space and ROM at the stabilized segment (P<0.05), but no significant changes were seen at the adjacent segments (P>0.05). X-ray scan showed neither instability or internal fixation loosen, breakage or distortion in follow-up. Conclusion: Dynesys system in addition to transfacet decompression through Wiltse approach is a therapy option for mild lumbar degenerative disease.This method can retention the structure of lumbar posterior complex and the activity of the fixed segment, reduce the risk of low back pain together with nerve root decompressed.The early clinical results are satisfactory.

[Effects and its mechanism of Nimotuzumab on radiosensitivity of esophageal carcinoma ECA-109 and TE-13 cell lines].

Objective: To investigate the effects of nimotuzumab on radiosensitivity of ECA-109 and TE-13 esophageal carcinoma cell lines and explore its possible mechanism. Methods: The ECA-109 and TE-13 cells were divided into control group, irradiation group, medicine group, and combined group (irradiation + medicine). In the combined group, ECA-109 and TE-13 cells were treated with nimotuzumab for 24 h before irradiation, and the cells were collected 2 h after irradiation. The radiosensitizing effects of nimotuzumab on ECA-109 and TE-13 cells were evaluated by clone formation assay. Cell apoptosis was detected by flow cytometry. Western blotting was used to evaluate the expression of EGFR, p-EGFR, DNA-PKcs, p-DNA-PKcs and γH2AX. Results: The values of Dq (quasithreshold dose), D0(mean lethal dose)and SF2 (surviving fraction at 2 Gy) of ECA-109 and TE-13 cells in the combined group were significantly lower than those of the radiation group (for ECA-109 cells, 1.11 vs. 1.72, 1.40 vs. 2.14, 0.42 vs. 0.66, respectively; for TE-13 cells, 0.41 vs. 0.46, 0.43 vs. 0.65, 0.40 vs. 0.71, respectively (all P<0.05). The sensitivity enhancement ratio (SER) of ECA-109 and TE-13 cells were 1.35 and 1.43, respectively. Flow cytometry showed that the apoptosis rate of ECA-109 and TE-13 cells in the combined group were significantly higher than those of the radiation group [for ECA-109 cells, (41.31±1.52)% vs. (9.54±0.52)%; for TE-13 cells, (46.28±0.28)% vs. (11.32±0.31)%, both P<0.01]. Western blotting showed that the expression levels of EGFR and DNA-PKcs were not significantly different in all groups (all P>0.05). Compared with those of the control group, p-EGFR and p-DNA-PKcs of the radiation group were significantly higher in both cell lines (P<0.05), and the γH2AX levels in the radiation group and medicine group were significantly higher than that of the control group (P<0.05). Compared with those of the radiation group and medicine group, p-EGFR and p-DNA-PKcs protein expression in the combined group were decreased significantly (P<0.05), while γH2AX protein expression was significantly increased (P<0.05). Conclusions: Nimotuzumab can enhance the radiosensitivity of esophageal cancer ECA-109 and TE-13 cells. The potential mechanism may be related to the inhibition of EGFR phosphorylation and down-regulation of DNA damage repair proteins. The radiosensitizing effect of nimotuzumab is greater on poorly differentiated esophageal cancer cells.

[Clinical features and gene mutations in epilepsy of infancy with migrating focal seizures].

Objective: To summarize the clinical features and gene mutations of epilepsy of infancy with migrating focal seizures (EIMFS). Method: Clinical features and electroencephalograms(EEG)of 9 patients with EIMFS of Peking University First Hospital from May 2015 to January 2016 were analyzed. Candidate gene mutations were screened by next generation sequencing. Result: Among the 9 patients, 3 were males and 6 were females. Two patients had family history. Seizure onset age was 2 days to 3 months after birth (median age 35 days). Migrating focal seizure was presented. Seizures manifested as eyes and(or)head deviation, involuntary blinking, swallowing, trembling or stiffness of limbs, hand clenching, flushing and cyanosis of lips, etc. Four patients had a history of status epilepticus. All 9 patients had psychomotor delay. EEG of all patients presented relatively slow background; during interictal phase, there were multi-focal epileptic discharges, which dominated one hemisphere or brain region; seizures were recorded in all 9 cases, which manifested eyes or(and)head deviation, stiffening or trembling of limbs, lip smacking, etc. Corresponding EEG showed low-medium-amplitude fast waves that originated from some brain regions and migrated to other regions. Cranial magnetic resonance imaging (MRI) was abnormal in 4 cases, which predominantly showed white matter dysplasia and enlargement of subarachnoid spaces. Two cases carried heterozygous missense mutations of SCN1A gene, while 3 cases carried heterozygous missense mutations of KCNT1 gene, all of which were de novo. One case carried compound heterozygous mutation of TBC1D24 gene(p.Gln207*, p. Ala289Va). Gene mutation was not found in 3 cases. All patients used multiple antiepileptic drugs (AED) and their seizures were not controlled. Follow-up ranged from 2 months to 5 years and 8 months, during which 4 were found dead. Two were lost to follow-up. Conclusion: EIMFS is clinically characterized by early onset, which is usually within 3 months after birth, migrating focal seizures, psychomotor delay, bad response to AED and high death rate. The interictal EEG showed multi-focal discharges, while ictal EEG shows migrating multifocal discharges. Genetic analysis can assist in diagnosis and genetic counseling.

[The medium conditioned incubation with rat heart cells maintains the properties of ES cells].

17 kinds of conditioned media were selected by plating tests, propagation and other tests with two cell lines, C-19-2 and MESPU-13. The results suggested that the rat heart cells conditioned media (RH-CM) can inhibit the spontaneous differentiation effectively, maintain the normal diploid karyotypes and promote adherence and proliferation of mouse ES cells. The ES cells were propagated in RH-CM to the 20th passage remaining their pluripotent in vivo and in vitro differentiation ability. RH-CM can be used as supplement of ES cell media. ES cells which are cultured in media containing 70% RH-CM and on PMEF feeder can maintain their undifferentiation state and diploid karyotype. RT-PCR detection suggested that there was mLIF expression in rat heart cells.

[The specific labelling of mouse embryonic stem cells].

Despite the wide application of ES cell technology, little is known about the pluripotent nature of ES cells. This is partly due to the heterogeneity of ES cell population in culture. This report described the specific labelling of undifferentiated cells in ES cell lines. oct-4 gene is specifically expressed in all the totipotent cells in mouse embryos and undifferentiated ES cells. A constructed pG18NG was obtained by inserting the reporter gene beta geo into the oct-4 transcription elements. ES cell lines MESPU22 and MESPU13 were transfected with this construct and stable integrated cell clones were selected out. With the experiments of in vitro cultivation, differentiation and chimeras production, it was confirmed that we have successfully labelled the undifferentiated cells in ES cell lines, and this label was both valid in vitro and in vivo.

[Establishment of two ES cell lines with good germline contribution].

Nine embryonic stem cell lines have been established from mouse strain 129/ter. Three of the nine ES cell lines were karyotypically normal. The nine cell lines showed some difference in the growth rate and the differential competence. Chimeras were made by injecting the ES cells into C57BL/6J blastocysts, and the germline compositions of the chimeras were detected by mating them with albino ICR mice. The results indicated that ES cell line MESPU21 and MESPU22 were both highly germline-competent. Comparing with other ones, these two cell lines both were karyotypically normal and propagated fast. The tissue composition of the teratocarcinomas derived from these cell lines appeared paralle to the results of chimera production. Careful manipulation during the process of ES cell establishment was helpful to obtain good cell lines.

[Formation of germline chimeras from murine embryonic stem cell lines].

Generation of germline chimeras is the crucial step in ES cell-mediated transgenesis. The prerequisite for germline chimerism is the maintenance of germline differentiating potency of ES cells, whereas production of germline chimeras is the only method to prove whether such potency is maintained. In order to investigate the germline differentiating potency of three newly established ES cell lines (MESPU21, MESPU22 and MESPU29), ES cells were introduced into host embryos from inbred C57BL/6J and outbred KMW or ICR through blastocyst injection or 8-cell stage morula injection. Totally 81 chimeras were obtained; among 42 test-bred ones, 19 were germline transmitters assessed by coat analysis, as is the first report of ES cell-embryo germline chimeras in China. MESPU21 and MESPU22 formed germline chimeras in high frequency and most of those chimeras produced ES cell-derived progeny in high proportion, which proved that both ES cell lines retained good germline differentiating potency and could be used as valuable cellular vehicles to introduce genetic modifications into mouse genome.

[Establishment of a high germline competent C57BL/6J ES cell line].

Eleven ES cell lines from C57BL/6J mice were established, with the primary culture of mice embryos as feeder cells and 1 x 10(3) units Leukemia Inhibitory Factor (LIF) in the DMEM (high glucose) medium. The frequency of establishment was 9.6%. Five of these ES cell lines were demonstrated karyotypically normal with diploid composition of over 70%. They shared the characteristics of early embryonic cells: positive for alkaline phosphatase activity and oct4 gene expression. They also showed high potency to differentiate into wide types of cells in vivo. Following injection of the blastocysts, three of them showed abilities to give rise to chimeras and MESPU35 cell line was identified to have high efficiency of colonization into the germline. The clones of MESPU35 cells maintained the germline competence and the mutant clone also retained the high potency to participate into the development of embryos. MESPU35 cells can serve as a valuable vehicle for the production of mutant mice.

Effects of calmodulin and calmodulin binding protein BP-10 on phosphorylation of thylakoid membrane protein.

Light-induced phosphorylation of endogenous thylakoid membrane protein can be inhibited markedly by a novel inhibitor CaMBP-10 which is discovered and isolated from plant. The inhibitory effect of BP-10 can be eliminated by addition of CaM. At the same time, the phosphorylation can also be inhibited by EGTA or CaM antagonists, such as TFP (trifluoperazine) and W-7 (N-(6-aminohexyl)-5-chloro-l-naphthalene sulfonamide). This result implies that (i) Ca(2+) and CaM most likely participate in and regulate plant photosynthesis; (ii) the kinase that catalyzes thylakoid membrane protein phosphorylation can be regulated by Ca(2+) and CaM. However, the further experiments indicate that BP-10 has no effect on dephosphorylation of thylakoid phosphoproteins.

[The analysis of GPI in chimeric mice of ES cells (MESPU 13)].

To estimate the differential potentiality of ES (Embryonic Stem) cell line MESPU 13, the heart, liver, spleen, lung, kidney, pancreas, gonad, muscle and blood of 19 chimeric mice were analyzed for GPI (Glucose Phosphate Isomeras) marker, in these studies, type A band from the ES cells, appeared parallel to the coat color chimerism of the mice. When coat color chimerism is below 40%, type A band was not seen except in the kidney of a few mice. Type A band was detected in nearly all the organs and tissues, when coat color chimerism was over 85%, indicated the ES cell has a fairly high potential to differentiate into cells of endoderm, mesoderm and ectoderm. In addition, only type A band was observed in the muscles of 6 mice which coat color chimerism is over 85%, the results indicated that these muscles were differentiated only from ES cells.

[Formation of mouse chimeras from early embryonic pluripotential stem cell].

Three chimeric mice were produced by injecting the embryonic stem cell-CCE cells into the cavity of 3.5 day host blastocysts from Kunming and C57BL/6J mice. By analyses of glucose phosphate isomerase (GPI-1), the results show a contribution from CCE cells in heart tissue of female Kunming chimera. Breeding experiments demonstrated that all chimeric mice were fertile but none of 3 chimeras produced germ line transmission.

[Effects of several primary factors related to establishment of mouse embryo pluripotent stem cell lines (ES cell line)].

Several primary factors related to establishment of mouse embryo pluripotent stem cell lines (ES cell lines) have been tested comparatively. The results of 825 embryos indicated: (1) Delayed blastocysts were more conducive than normal ones to proliferation of inner cell mass (ICM). (2) Culture medium DMEM with 4 500 mg/l glucose was more advantageous than DMEM with 1000 mg/l glucose to the attachment of explanted blastocysts and to the proliferation of ICM. (3) DMEM without sodium pyruvate was beneficial to appearance of ES cell colonies and to their growth. (4) Feeder layer prepared from the primary culture of mouse embryos had very good effects to the growth and culture of ES cells.

[Preparation and utilization of BrdU antiserum. II. Detections of BrdU labelling chromosome and DNA using BrdU antibody technique].

Utilizing highly specific BrdU antiserum, via the second antibody conjugated with enzyme and DAB-H2O2 or AEC-H2O2 colorific methods, the BrdU marked regions of CHO and human chromosomes were detected clearly. The BrdU-DNA prints (pg) on nitrocellulose membranes were also showed with similar methods.

[Enzyme-linked immunosorbent assay of larvicidal toxic proteins of Bacillus sphaericus Ts-1].

Bacillus sphaericus Ts-1 Mosquito larvicidal toxins 42 k Da and 43 k Da were isolated by Sephadex G-200 chromatography. Three strains of highly toxic B. sphaericus and two non toxic strains were screened for toxic proteins using ELISA. The lowest detectable toxin level was 1.56 X 10(-5) mg/ml. Non toxic strains did not produce antigens reacting to either the 42 kDa or the 43 kDa antibodies. Ts-1 cultures were examined at 12 and 24 h by LC50 bioassay against Culex pipiens. The LC50's at 12 h and 24 h were 0.71 ppm and 0.154 ppm, respectively, i.e., the toxin level at 24 h was 4.6 times the level at 12 h. ELISA tests established total toxin at 0.049 mg/ml and 0.225 mg/ml at 12 h and 24 h, respectively, confirming the LC50 study.

[Separation and purification of the toxic protein of Bacillus sphaericus Ts-1].

Bacillus sphaericus strain Ts-1 is highly insecticidal to larvae of the mosquito. It's insecticidal component is toxic proteins. The toxin was extracted from spore-crystal complexes by disruption in a Sonicator Cell Disruptor Model W-220F followed by treatment with 0.05 mol/L NaOH. Fraction recovered from chromatography of the spore-crystal complexes on column of Sephadex G-200 were assayed against mosquito larvae and the toxic fractions from gel chromatography were subjected to SDS-PAGE. The toxic proteins in B. sphaericus Ts-1 spore-crystal complex migrated in position corresponding to 42kD and 43kD. Bioassay of the two purified proteins prepared by PAGE indicated that they were all toxic to mosquito larvae. Toxic protein was further purified by DEAE-cellulose chromatography. The toxic protein with a molecular weight of 42kD was obtained.