Multiplexed Representation of Itch and Mechanical and Thermal Sensation in the Primary Somatosensory Cortex.

The primary somatosensory cortex (S1) plays a critical role in processing multiple somatosensations, but the mechanism underlying the representation of different submodalities of somatosensation in S1 remains unclear. Using in vivo two-photon calcium imaging that simultaneously monitors hundreds of layer 2/3 pyramidal S1 neurons of awake male mice, we examined neuronal responses triggered by mechanical, thermal, or pruritic stimuli. We found that mechanical, thermal, and pruritic stimuli activated largely overlapping neuronal populations in the same somatotopic S1 subregion. Population decoding analysis revealed that the local neuronal population in S1 encoded sufficient information to distinguish different somatosensory submodalities. Although multimodal S1 neurons responding to multiple types of stimuli exhibited no spatial clustering, S1 neurons preferring mechanical and thermal stimuli tended to show local clustering. These findings demonstrated the coding scheme of different submodalities of somatosensation in S1, paving the way for a deeper understanding of the processing and integration of multimodal somatosensory information in the cortex.SIGNIFICANCE STATEMENT Cortical processing of somatosensory information is one of the most fundamental aspects in cognitive neuroscience. Previous studies mainly focused on mechanical sensory processing within the rodent whisking system, but mechanisms underlying the coding of multiple somatosensations remain largely unknown. In this study, we examined the representation of mechanical, thermal, and pruritic stimuli in S1 by in vivo two-photon calcium imaging of awake mice. We revealed a multiplexed representation for multiple somatosensory stimuli in S1 and demonstrated that the activity of a small population of S1 neurons is capable of decoding different somatosensory submodalities. Our results elucidate the coding mechanism for multiple somatosensations in S1 and provide new insights that improve the present understanding of how the brain processes multimodal sensory information.

Itch perception is reflected by neuronal ignition in the primary somatosensory cortex.

Multiple cortical areas including the primary somatosensory cortex (S1) are activated during itch signal processing, yet cortical representation of itch perception remains unknown. Using novel miniature two-photon microscopic imaging in free-moving mice, we investigated the coding of itch perception in S1. We found that pharmacological inactivation of S1 abolished itch-induced scratching behavior, and the itch-induced scratching behavior could be well predicted by the activity of a fraction of layer 2/3 pyramidal neurons, suggesting that a subpopulation of S1 pyramidal neurons encoded itch perception, as indicated by immediate subsequent scratching behaviors. With a newly established optogenetics-based paradigm that allows precisely controlled pruritic stimulation, we found that a small fraction of S1 neurons exhibited an ignition-like pattern at the detection threshold of itch perception. Our study revealed the neural mechanism underlying itch perceptual coding in S1, thus paving the way for the study of cortical representation of itch perception at the single-neuron level in freely moving animals.

Pediatric acute myeloid leukemia patients with i(17)(q10) mimicking acute promyelocytic leukemia: Two case reports.

BACKGROUND: Chromosome i(17)(q10) abnormality is mainly associated with chronic myeloid leukemia (CML), myelodysplastic syndrome/myeloproliferative tumors (MDS/MPD), and acute myeloid leukemia (AML). The role of i(17)(q10) in AML is still unknown, the differences between AML and acute promyelocytic leukemia (APL)-like AML with i(17)(q10) need more research. This study aimed to investigate the clinical characteristics and laboratory evidence of 2 AML cases with i(17)(q10), similar to APL phenotype. CASE SUMMARY: Both pediatric patients were males; case 1 had newly diagnosed AML, and case 2 showed relapsed tumor after 1 year of drug withdrawal. Bone marrow cell morphology, chromosome karyotype analysis, Fully-instrumented submersible housing test, immunological assays, molecular biological methods, and blood tumor panoramic gene test were performed. All-trans retinoic acid (ATRA) combined with arsenic acid (As2O3) were used in the first course of treatment. Bone marrow was dominated by abnormal promyelocytic granulocytes. Karyotype test revealed i(17)(q10) isochromosome. Immunological phenotype mainly included positive expressions of CD9, CD13, CD33, and CD38. Case 1 suffered intracranial hemorrhage after re-chemotherapy and died on D162. For case 2, on D145 and D265, bone marrow promyelocytic granulocytes accounted for 2%. Flow cytometric residual lesion detection showed no abnormal immunophenotype cells. The copy number of WT1 gene in two cases were 1087 and 1010, respectively, and the expression rates were 55.29% and 59.5%, respectively. CONCLUSION: ATRA, As2O3, and chemotherapy may be ineffective in treating APL-like AML with i(17)(q10) but without t(15;17) and PML-RARA fusion gene.

[Effect of human follicular fluid from endometriosis patients treated with quyu jiedu granule on mouse embryonic development].

OBJECTIVE: To observe the effect of human follicular fluid (HFF) from endometriosis (EM) patients treated with Quyu Jiedu Granule (QJG) on mouse embryonic development. METHODS: Cultured 2-cell mouse embryos were divided into three groups. To the medium of Group A (70 embryos), HFF from endometriosis patients treated with QJG was added; to that of Group B (60 embryos), HFF from endometriosis patients untreated with QJG, and to Group C (59 embryos), HFF from patients with fallopian tube obstruction was added. The percentages of embryos developed to 8-cell stage, morula stage and blastula stage were counted, and the early stage number and rate of high-quality embryo were measured. RESULTS: Among the 70 embryos in Group A, 53 (75.71%) developed to 8-cell stage, 48 (68.57%) to morula stage and 45 (64.28%) to blastula stage; while in the 60 embryos of Group B, the corresponding number (percentage) were 34 (56.67%), 29 (48.33%), 21 (35.00%), respectively, showing significant differences between groups (P < 0.05). High-quality of embryo was 61 (87.14%) in Group A and 44 (73.3%) in Group B, the difference between groups also showed statistical significance (P < 0.05). CONCLUSION: HFF from endometriosis patients is toxic to 2-cell mouse embryos, but after treated by QJG, it could elevate the quality of cultured mouse embryo in the early stage.

[Effects of Quyu Jiedu Granule on expressions of tumor necrosis factor-alpha and interleukin-6 mRNAs in ovarian granulosa cells of endometriosis rats].

OBJECTIVE: To observe the effects of Quyu Jiedu Granule, a compound traditional Chinese herbal medicine for removing blood stasis and expelling superficial evils, on the quality of oocytes and the expressions of tumor necrosis factor-alpha (TNF-alpha)and interleukin-6 (IL-6) mRNAs in ovarian granulosa cells of endometriosis (EM) rats. METHODS: Forty EM rats were randomly divided into two groups: experimental group and control group. There were 20 EM rats in each group. The uteri of another 20 SD rats were drawn in sham-operated group. The number and percentage of high quality oocytes and the levels of TNF-alpha and IL-6 mRNA expressions in the granulosa cells of EM rats were detected by using reverse transcription-polymerase chain reaction. RESULTS: The number and percentage of high quality oocytes in the experimental group were significantly higher than those in the control group (P<0.05), and the levels of TNF-alpha and IL-6 mRNAs in granulose cells in the experimental group were significantly lower than those in the control group (P<0.05). CONCLUSION: The increase of the TNF-alpha and IL-6 mRNA expressions in ovarian granulose cells of EM rats leads to the decrease of the oocyte quality. The mechanism of Quyu Jiedu Granule in improving the quality of oocytes may be related to the decrease of TNF-alpha and IL-6 mRNA expressions in ovarian granulosa cells.

Expressions of VEGF and Ki-67 in eutopic endometrium of patients with endometriosis and effect of Quyu Jiedu Recipe on VEGF expression.

OBJECTIVE: To observe the clinical efficacy of Quyu Jiedu Recipe (, QJR) in treating endometriosis (EM), and to explore the levels of vascular endothelial growth factor (VEGF) and cell proliferative nucleoprotein antigen (Ki-67), their changes before and after treatment and the clinical significance in the trial. METHODS: Fifty patients of EM were randomly assigned to two groups. The 26 patients in the QJR group were treated with QJR, and the 24 patients in the gestrinone (GT) group with gestrinone. Besides, a normal control group with 20 healthy women was set up. The therapeutic effects in the two treated groups were compared. Expressions of VEGF and Ki-67 in eutopic endometrium of all subjects (with both patients and healthy women at the median secretive phase) were determined with immunohistochemical stain before treatment, and the determination in the two treated groups was repeated after 3-month treatment in the same phase. RESULTS: Before treatment, the VEGF and Ki-67 expression positive rates and their mean optic density (MOD) were higher in patients than in healthy women (P<0.05). After treatment, the positive rate and MOD of VEGF expression lowered significantly than before treatment (P<0.05), but those of Ki-67 changed insignificantly, and comparison between the two treated groups showed no significant difference (P>0.05). CONCLUSION: QJR could markedly improve the symptoms of menorrhagia and menstrual disorder, and its mechanism might be related with the lowering of eutopic endometrial VEGF expression. VEGF and Ki-67 show a high expression in eutopic endometrium of patients with EM.

Characterization of ORF39 from Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus, the gene containing RNA recognition motif.

In the genome of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus, open reading frame 39 (Ha39) is the only gene predicted to encode an RNA recognition protein. Computer analysis revealed that Ha39 homologues were found in 15 NPVs, but not in GVs. Its transcripts were detected from 3 through 72 hours post infection (h p.i.) using RT-PCR and Northern blot analysis. The protein was detected in infected-cell lysates from 6 h p.i. Western blot assay of ODV and BV preparations revealed that Ha39 encodes a structural protein associated with BVs.Additionally, immunofluorescence microscopy demonstrated that the protein was present within cytoplasm in virusinfected cells, but not in the nuclear region.

Characterization of Ha29, a specific gene for Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus.

Open reading frame 29 (ha29) is a gene specific for Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearSNPV). Sequence analyses showed that the transcription factor Tfb2 motif, bromodomain and Half-A-TPR (HAT) repeat were present at aa 66-82, 4-76, 55-90 of the Ha29 protein respectively. The product of Ha29 was detected in HearSNPV-infected HzAM1 cells at 3 h post-infection. Western blot analysis using a polyclonal antibody produced by immunizing a rabbit with purified GST-Ha29 fusion protein indicates that Ha29 is an early gene. The size of Ha29 product in infected HzAM1 cells was about 25 kDa, which was larger than the presumed size of 20.4 kDa. Tunicamycin treatment of HearSNPV-infected HzAM1 cells suggested that the Ha29 protein is N-glycosylated. Fluorescent confocal laser scanning microscope examination, and Western blot analysis of purified budded virus (BVs), occlusion-derived virus (ODVs), cell nuclear and cytoplasmic fraction, showed that the Ha29 protein was localized in the nucleus. Our results suggested that ha29 of HearSNPV encodes a non-structurally functional protein that may be associated with virus gene transcription in Helicoverpa hosts.

[Research progress in malate dehydrogenase(MDH) of honeybees].

Malate dehydrogenase(MDH) is an important enzyme in glycometabolism. MDH of Apis mellifera showed three enzyme active zones, MDHI,MDHIIand MDHIII. MDHIand MDHIII maintained relative stability in different castes and developmental phases,but MDHIIwas polymorphic,and controlled by three alleles,a,b and c.MDH of Apis cerana was coded by S and F alleles,but some authors reported it is monomorphic.MDH was applied to the studies of A. mellifera, which included several aspects as follows: the number of queen matings,labor division in honeybee societies, the analysis of genetic constitution in honeybee populations and so on. The combination of both MDH and molecular biology will certainly promote honeybee studies.

Effect of Quyu Jiedu granule on microenvironment of ova in patients with endometriosis.

OBJECTIVE: To observe the effect of Quyu Jiedu Granules (, QJG) on the micro- microenvironment of ova in patients with endometriosis (EM). environment METHODS: Twenty EM patients who received in vitro fertilization and embryo transfer (IVF-ET) were randomized equally into a treated group and a control group. Further, 20 patients who received IVF-ET due to oviduct factors were enrolled into a non-endometriosis group. The dosage of gonadotrophic hormone used, the number of ova attained, fertilization rate and clinical pregnancy rate were all observed, and the levels of tumor necrosis factor alpha (TNF-right harpoon over left harpoon) and interleukin 6 (IL-6) in follicular fluid as well as their mRNA expressions in ovarian granular cells were detected by RT-PCR on the very day of ovum attainment. RESULTS: The ova attainment (13.80+/-6.87) and fertilization rate (0.69+/-0.31) in the treated group were all higher than the corresponding values in the control group (9.80+/-5.32 and 0.47+/-0.22); the follicular fluid contents of TNF-alpha and IL-6 in the treated group were 1.38+/-0.21 ng/mL and 130.56+/-12.81 pg/mL, respectively, which were lower than those in the control group (1.98+/-0.34 ng/mL and 146.83+/-17.65 pg/mL, respectively). Further, the treated group showed much lower mRNA expressions of TNF-alpha and IL-6 in ovarian granular cells. CONCLUSIONS: The elevation of TNF-alpha and IL-6 contents in follicular fluid and their mRNA expressions in ovarian granular cells are possibly related to the low quality of ova in EM; QJG might raise the ova quality by reducing TNF-alpha and IL-6 levels to improve the living micro-environment for the ova.

Characterization of a unique gene ORF135 from Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus.

The ORF135 of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearSNPV)(Ha135) is one of the 20 genes that are unique to HearSNPV. Computer-assisted analysis revealed that four potential post translation modification sites, four transcription factor associated domains and a DNA binding protein domain were found in Ha135 amino acid sequence. Northern blot analysis of Ha135 indicated that Ha135 transcript was detected at 12 h.p.i. and remained detectable at up to 122 h.p.i. RT-PCR method was used to understand the temporal regulation of the transcript at earlier stages, the result showed that the Ha135 transcript was detected as early as 3 h p.i. suggesting that Ha135 was an early gene, which is in agreement with the early promoter motifs. The Ha135 protein was also detected at 12 h.p.i and remained detectable until 122 h.p.i. by western blot using an anti-Ha135 antiserum. The product of Ha135 was found to be about 29 kDa, bigger than the predicted 24 kDa molecular weight, suggesting that post translational modification of the Ha135 protein occur in host cells. The subcellular location was studied using EGFP-Ha135, which suggested that the Ha135 protein is primarily localized in the nucleus, which is compatible with several functional domains present in Ha135 amino acid sequence. Together, these results suggest the possibility that HearSNPV ORFI35 might be involved in viral DNA transcription and/or replication.