Map-based cloning and functional analysis revealed ABCC2 is responsible for Cry1Ac toxin resistance in Bombyx mori.

Bt toxins are parasporal crystals produced by Bacillus thuringiensis (Bt). They have specific killing activity against various insects and have been widely used to control agricultural pests. However, their widespread use has developed the resistance of many target insects. To maintain the sustainable use of Bt products, the resistance mechanism of insects to Bt toxins must be fully clarified. In this study, Bt-resistant and Bt-susceptible silkworm strains were used to construct genetic populations, and the genetic pattern of silkworm resistance to Cry1Ac toxin was determined. Sequence-tagged site molecular marker technology was used to finely map the resistance gene and to draw a molecular genetic linkage map, and the two closest markers were T1590 and T1581, indicating the resistance gene located in the 155 kb genetic region. After analyzing the sequence of the predicted gene in the genetic region, an ATP binding cassette transporter (ABCC2) was identified as the candidate gene. Molecular modeling and protein-protein docking result showed that a tyrosine insertion in the mutant ABCC2 might be responsible for the interaction between Cry1Ac and ABCC2. Moreover, CRISPR/Cas9-mediated genome editing technology was used to knockout ABCC2 gene. The homozygous mutant ABCC2 silkworm was resistant to Cry1Ac toxin, which indicated ABCC2 is the key gene that controls silkworm resistance to Cry1Ac toxin. The results have laid the foundation for elucidating the molecular resistance mechanism of silkworms to Cry1Ac toxin and could provide a theoretical basis for the biological control of lepidopteran pests.

[Clinical features of children with myelin oligodendrocyte glycoprotein antibody-associated disorders].

OBJECTIVE: To study the clinical features and treatment outcome of children with myelin oligodendrocyte glycoprotein (MOG) antibody-associated disorders (MOGAD). METHODS: A retrospective analysis was performed for the clinical data of 28 children with MOGAD (with 38 demyelinating episodes). RESULTS: Among the disease spectrums of 28 children with MOGAD, optic neuritis was the most common (12 cases, 43%), followed by acute disseminated encephalomyelitis (9 cases, 32%). Among the 38 demyelinating episodes in the 28 children, there were 29 cases (76%) of lesions in the acute stage on head magnetic resonance imaging (MRI), and most of these lesions were extensive or isolated subcortical white matter lesions. A total of 24 cases of spinal MRI results in the acute stage were recorded, among which there were 11 cases (46%) of spinal lesions. MRI abnormalities of the optic nerve were found in 18 cases of optic neuritis in the acute stage. Of the 28 children, 20 (71%) had an increase in white blood cell count in cerebrospinal fluid, with lymphocytes as the most common type of cells, and 3 children had an increase in protein. The titer of serum MOG antibody was 1:10-1:320 in the 28 children. All 28 children were administered with glucocorticoids, along with immunoglobulin in 18 children. The symptoms of 26 children (93%) were alleviated during follow-up, and only 2 children had neurological sequela of the optic function. CONCLUSIONS: The clinical manifestations are diverse in children with MOGAD. Immunotherapy is effective and most children have a good prognosis.

[A comparative analysis of anti-N-methyl-D-aspartate receptor encephalitis with or without abnormal findings on cranial magnetic resonance imaging].

OBJECTIVE: To investigate the clinical features of children with anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis with normal or abnormal cranial magnetic resonance imaging (MRI) findings via a comparative analysis. METHODS: A retrospective analysis was performed for the clinical data of 33 children with anti-NMDAR encephalitis. The clinical features and prognosis were compared between the children with normal and abnormal cranial MRI findings. RESULTS: In the 33 children with anti-NMDAR encephalitis, the most common initial symptoms were seizures (61%) and involuntary movement (61%), followed by language disorder (54%), mental and behavioral abnormalities (52%), and disturbance of consciousness (30%). All children had positive anti-NMDAR antibody in the cerebrospinal fluid, and 29 children (88%) had positive serum antibody. Of all the children, 15 (46%) had increased leukocytes in the cerebrospinal fluid, 3 (9%) had an increase in protein, and 29 (88%) had positive oligoclonal band; 26 children (79%) had electroencephalographic abnormalities (epileptic wave, slow wave, or a combination of these two types of waves). One child experienced respiratory failure. One child was found to have germinoma in the sellar region during follow-up. Of all the 33 children, 13 (39%) had abnormal cranial MRI findings, with hypointensity or isointensity on T1W1 and hyperintensity on T2WI and T2-FLAIR; 2 children had dural enhancement. As for the location of lesion, 5 children (38%) had lesions in the temporal lobe, 3 (23%) in the frontal lobe, 3 (23%) in the basal ganglia, 2 (15%) in the parietal lobe, 2 (15%) in the occipital lobe, 2 (15%) in the brainstem, 1 (8%) in the thalamus, and 1 (8%) in the cerebellum. Among the 13 children with abnormal cranial MRI findings, 5 (38%) had lesions mainly in the grey matter and 8 (62%) had lesions mainly in the white matter. Compared with the children with normal cranial MRI findings, the children with abnormal cranial MRI findings had significantly higher proportion of children with prodromal infection, incidence rate of disturbance of consciousness, probability of recurrence, Glasgow score, incidence rate of increased leukocytes in the cerebrospinal fluid, and application rate of second-line treatment (P<0.05). CONCLUSIONS: Children with anti-NMDAR encephalitis and abnormal cranial MRI findings have certain clinical features, which may provide guidance for the evaluation of disease conditions and the selection of diagnostic and treatment measures.

LncRNA CDKN2B­AS1 sponges miR­28­5p to regulate proliferation and inhibit apoptosis in colorectal cancer.

Long noncoding RNA (lncRNA) CDKN2B­antisense RNA 1 (AS1) functions as a tumor oncogene in numerous cancers. However, the roles and mechanism of CDKN2B­AS1 in colorectal cancer (CRC) have not been explored. The present study aimed to investigate whether and how CDKN2B­AS1 contributes to CRC progression. The data revealed that CDKN2B­AS1 expression was upregulated in CRC tissues. Loss­of­function assays demonstrated that CDKN2B­AS1 in CRC modulated cell proliferation and apoptosis, which was mediated by cyclin D1, cyclin­dependent kinase (CDK) 4, p­Rb, caspase­9 and caspase­3. Bioinformatics analysis and luciferase reporter assays indicated direct binding of microRNA (miR)­28­5p to CDKN2B­AS1. Moreover, the results herein revealed that the expression of miR­28­5p was negatively correlated with that of CDKN2B­AS1 in CRC tissue. Moreover, CDKN2B­AS1 acted as a miR­28­5p competing endogenous RNA (ceRNA) to target and regulate the expression of URGCP. These findings indicated that CDKN2B­AS1 plays roles in CRC progression, providing a potential therapeutic target or novel diagnostic biomarker for CRC.

Fractionated resection on low grade gliomas involving Broca's area and insights to brain plasticity.

BACKGROUND: Recent advances on functional mapping have enabled us to conduct surgery on gliomas within the eloquent area. The objective of the article is to discuss the feasibility of a planned fractionated strategy of resection on low-grade gliomas (LGGs) involving Broca's area. We report the first surgical series of planned fractionated resections on LGGs within Broca's area, focusing on language functional reshaping. METHODS: Four patients were treated with fractionated operations for LGGs involving Broca's area. All cases underwent conventional magnetic resonance (MR) scanning, language functional MR and diffusion tensor imaging (DTI) before operation. The resections were then performed on patients under awake anesthesia using intraoperative electrical stimulation (IES) for functional mapping. Pre- and post-operative neuro-psychological examinations were evaluated. RESULTS: Total resections were achieved in all cases as confirmed by the postoperative control MR. After transient language worsening, all patients recovered to normal 3-6 months later. Language functional MR scannings have shown language functional cortical and subcortical pathway reorganization (in the perilesion or contra-lateral hemisphere) after the operation. All patients returned to a normal socioprofessional life. CONCLUSIONS: By utilizing the dynamic interaction between brain plasticity and fractionated resections, we can totally remove the tumor involving Broca's structure without inducing permanent postoperative deficits and even improve the quality of life.

[Analysis of DNMT3a mutation in childhood acute myeloid leukemia].

Within the past few years, the invention of next-generation sequencing has revealed several new genes associated with tumor formation and development, for example DNMT3a. This gene is an independent prognostic factor for acute myeloid leukemia (AML). The objective of this study was to analyze the DNMT3a mutation in childhood AML in a single center. PCR amplification of the entire coding region of DNMT3a was performed using 23 overlapping primer pairs in 57 patients who were diagnosed in Blood Disease Hospital of Chinese Academy of Medical Sciences, then the directly sequencing was underwent. The results showed that no DNMT3a mutation was found in these patients including the hotspot R882. But AML1/ETO mutation was found in 10 patients, CBFB/MYH11 mutation in 3 patients, PML/RARa mutation in 13 patients, FLT3/ITD mutation in 5 patients, FLT3/TKD mutation in 1 patient, PML/RARa and FLT3/TKD mutation coexisted in 2 patients. It is concluded that DNMT3a mutations are rare in childhood AML, and different mechanisms of myeloid leukemogenesis between childhood and adults maybe involved.

[Diagnostic value of fluorescence in situ hybridization for children with acute myeloid leukemia].

This study was purposed to investigate the diagnostic value of fluorescence in situ hybridization (FISH) technique for the childhood acute myeloid leukemia (AML). The medical data of 179 children with AML (aged ≤ 16 years) were retrospectively studied, who were initially diagnosed in our hospital from April 2005 to April 2010. Through the analysis of the results of FISH, chromosome banding analysis and polymerase chain reaction, the difference and complementarity between FISH and other 2 methods for detecting the fusion genes were explored. The results indicated that the detection rate of genetic abnormality with FISH was higher. The PML/RARα probe was used in 27 AML-M3 patients, 22 out of whom were evaluated as PML/RARα positive. The AML1/ETO probe was used in 24 AML-M2b patients and all of them were evaluated as AML1/ETO positive. The CBFß/MYH11 probe was used in 4 AML-M4Eo patients and all of them were evaluated as CBFß/MYH11 positive. It is concluded that FISH is a sensitive method for detecting fusion genes and the results of FISH have a good correlation with the chromosome banding analysis and polymerase chain reaction. The combination of FISH with other 2 methods improves the detection rate of genetic abnormality, which is useful for the diagnosis and typing of childhood AML.

[Large sample clinical analysis of patients with children acute leukemia in single center].

In order to investigate the epidemiology of childhood acute leukemia (CAL), such as onset age and time, risk factor, subtypes distribution and genetics, 1236 CAL patients admitted in blood disease hospital of Chinese Academy of Medical Sciences for treatment from April 2004 to April 2010 were analyzed retrospectively. The results showed that the sex ratio of ALL and AML patients were 1.80:1 and 1.73:1 respectively; the average peak age of incidence lasted from 2 to 6 years with the median age of 6 years, while the ALL peak age of incidence lasted from 2 to 5 years but AML showed no significant peak age of incidence. Winter, especially January was the peak time for both onset and birth. Among all the 631 ALL patients who had already been immunophenotyped, B-ALL patients accounted for 83%, T-ALL patients accounted for 9%. Among 361 AML patients, sub-leukemia phenotype from M(0) to M(7) accounted for 0.3%, 2.2%, 29.8%, 20.9%, 8.1%, 25.2%, 4.1% and 4.6% respectively. Among 631 pediatric ALL patients who had been examined by using molecular biology technique, the positive rate of TEL/AML1, BCR/ABL, MLL and E2A/PBX1 were 23%, 7.4%, 4.1%, 2.1% respectively. Among 361 pediatric AML patients who had been examined by using molecular biology technique, 19% of the patients showed positive AML1/ETO fusion gene, 18% of the patients showed positive PML/RARα fusion gene, while 4.2% of patients showed positive CBFß/MYH11. It is concluded that the onset of pediatric acute leukemia is influenced by age, season, environment and different genetic background.