Objective To promote the development of precision medicine and gene sequencing technologies in China by analyzing the innovation situation in gene sequencing technologies.Methods An innovation situation research analysis frame was established with scientific literature and patent information as its basic data.The innovation situation in gene sequencing technologies was analyzed in aspects of scientific research and technical R & D.Results The research on gene sequencing technologies and technical R & D were focused on tumor diagnosis and treatment,plant gene sequencing,identification of HIV and gastrointestinal flora.Single cell genome amplification technology has greatly promoted the development of gene sequencing technologies.Conclusion The level of gene sequencing technologies and technical R & D in China is approaching to that in the world.

Objective To investigate the effect of HbA1c meeting the standard or not on microalbuminuria,blood lipids and liver enzymes in patients with type 2 diabetes.Methods A retrospective analysis was performed on 457 subjects who had type 2 diabetes.They were divided into substandard group and standard group according to HbA1c result.The general information and relevant laboratory indicators of patients were.collected and compared between two groups.Results The microalbuminuria,serum triglyceride and liver enzymes (glutamyl transpeptidase,alkaline phosphatase,aspertate aminotransferase) were significantly different between two groups [ (189.8 ± 235.3) mg/dl vs (38.9 ± 85.5) mg/dl,(2.64 ± 2.99) mmol/L vs (2.02 ± 1.50)mmol/L,(41.7 ±52.9)U/L vs (29.7 ±24.9)U/L,(83.6 ±28.6) U/L vs (74.3 ±25.8)U/L,(26.7 ±19.1)U/L vs (22.0 ±10.5) U/L,P ＜0.05].HbA1c level was positively correlated with microalbuminuria,glutamyl transpeptidase and alkaline phosphatase (r =0.209,0.115,0.11,P ＜0.01).The microalbuminuria was an independent risk factor of affecting HbA1c to reach the standard (OR = 1.009,P ＜0.05).Conclusions HbA1c meeting the standard or not can influence many factors except blood glucose.

Objective A major component of flow cytometry data analysis involves gating , which is the process of identifying homogeneous groups of cells .As manual gating is error-prone, non-reproducible, nonstandardized, and time-consuming , we propose a time-efficient and accurate approach to automated analysis of flow cytometry data .Methods Unlike manual analysis that successively gates the data projected onto a two-dimensional filed, this approach, using the K-means clustering results , directly analyzed multidimensional flow cytometry data via a similar subpopulations-merged algorithm.In order to apply the K-means to analysis of flow cytometric data , kernel density estimation for selecting the initial number of clustering and k-d tree for optimizing efficiency were proposed .After K-means clustering , results closest to the true populations could be achieved via a two-segment line regression algorithm .Results The misclassification rate (MR) was 0.0736 and time was 2 s in Experiment One, but was 0.0805 and 1 s respectively in Experiment Two. Conclusion The approach we proposed is capable of a rapid and direct analysis of the multidimensional flow cytometry data with a lower misclassification rate compared to both nonprobabilistic and probabilistic clustering methods .

Objective:Hidden hunger remains a severe public health problem that affects millions of people worldwide.In China,challenges related to dietary imbalance and hidden hunger persist.Micronutrient inadequacy deserves more attention among adolescents,given its vital role in their growth and development;however,this problem appears to have been largely ignored.High school students,in particular,are often at a high risk of hidden hunger but have limited assessment tools available.Therefore,this study aims to revise the hidden hunger assessment scale for high school students(HHAS-HSS)in China and assess its reliability and validity.Methods:Based on a literature review,expert consultation,pre-experiment,and formal survey,a hidden hunger assessment scale was revised for high school students.The formal survey involved 9 336 high school students in 11 of the 16 cities in Anhui Province,China,and 9038 valid questionnaires were collected and included in the analysis.The item analysis,internal consistency reliability,test-retest reliability,content validity,exploratory factor analysis,and confirmatory factor analysis of the HHAS-HSS were examined.Results:The HHAS-HSS included a total of 4 dimensions and 12 items:"vegetables and food diversity"(three items),"fruits and dairy products"(three items),"micronutrient-dense foods"(four items),and"health condition and eating habits"(two items).The results showed a Cronbach's alpha of 0.758,a split-half reliability of 0.829,and a test-retest reliability of 0.793,indicating good internal consistency.Using the Bartlett's test and Kaiser-Meyer-Olkin test(KMO)to test the exploratory factor analysis presented a four-factor model of the HHAS-HSS,the KMO value was 0.820(P＜0.001),which indicated the possibility for factor confirmatory factor analysis.Using the maximum variance rotation method,four factors were obtained,and the cumulative variance explained rate was 57.974％.Confirmatory factor analysis also supported the division of the scale into four dimensions,and the fitting indices were x2=1417.656,x2/df=29.534,goodness-of-fit index=0.974,adjusted goodnesss-of-fit index=0.958,parsimonious goodness-of-fit index=0.600,normed fit index=0.938,incremental fit index=0.940,Tucker-Lewis index=0.917,comparative fit index=0.939,and root mean square error of approximation=0.056.Except for x2/df,all the indices reached the fitting standard,and the above results showed that the construct validity of the scale reached an acceptable level.Conclusions:The HHAS-HSS has good validity and reliability for Chinese high school students.It is a convenient self-report measure of hidden hunger risk.

5-Hydroxytryptamine (5-HT) type 3 receptor (5-HT₃R) is the only type of ligand-gated ion channel in the 5-HT receptor family. Through the high permeability of Na⁺, K⁺, and Ca²⁺ and activation of subsequent voltage-gated calcium channels (VGCCs), 5-HT₃R induces a rapid increase of neuronal excitability or the release of neurotransmitters from axon terminals in the central nervous system (CNS). 5-HT₃Rs are widely expressed in the medial prefrontal cortex (mPFC), amygdala (AMYG), hippocampus (HIP), periaqueductal gray (PAG), and other brain regions closely associated with anxiety reactions. They have a bidirectional regulatory effect on anxiety reactions by acting on different types of cells in different brain regions. 5-HT₃Rs mediate the activation of the cholecystokinin (CCK) system in the AMYG, and the γ‍-aminobutyric acid (GABA) "disinhibition" mechanism in the prelimbic area of the mPFC promotes anxiety by the activation of GABAergic intermediate inhibitory neurons (IINs). In contrast, a 5-HT₃R-induced GABA "disinhibition" mechanism in the infralimbic area of the mPFC and the ventral HIP produces anxiolytic effects. 5-HT₂R-mediated regulation of anxiety reactions are also activated by 5-HT₃R-activated 5-HT release in the HIP and PAG. This provides a theoretical basis for the treatment of anxiety disorders or the production of anxiolytic drugs by targeting 5-HT₃Rs. However, given the circuit specific modulation of 5-HT₃Rs on emotion, systemic use of 5-HT₃R agonism or antagonism alone seems unlikely to remedy anxiety, which deeply hinders the current clinical application of 5-HT₃R drugs. Therefore, the exploitation of circuit targeting methods or a combined drug strategy might be a useful developmental approach in the future.
Serotonin ; Receptors, Serotonin, 5-HT3 ; Anxiety ; Neurons ; gamma-Aminobutyric Acid

ObjectiveTo observe the effects of Cuscutae Semen on the learning and memory ability， N6-methyladenosine （m⁶A）-related modification enzymes and total m⁶A level in hippocampus of the offspring of fear-damaged pregnant rats. MethodForty-five pregnant rats were randomly divided into blank group， model group and Cuscutae Semen group. From the 1^st day to the 19^th day of pregnancy， rats in the model group and the Cuscutae Semen group were induced by observing electric shock of other rats. The Cuscutae Semen group was treated with 5 g·kg^-1·d^-1 Cuscutae Semen decoction （ig）， while the other two groups were treated with the same amount of purified water. The offspring were assigned following the grouping method of their maternal generation. The behavioral changes of the offspring were tested by Morris water maze on 21^st day after birth， and the development of hippocampal neurons was observed by transmission electron microscopy. The mRNA and protein expression levels of methyltransferase-like 3 （METTL3）， METTL14， Wilms tumor 1 associated protein （WTAP）， fat mass and obesity-associated protein （FTO） and Alk B homologue 5 （ALKBH5） were detected by Real-time polymerase chain reaction （Real-time PCR）， Western blot and immunohistochemistry （IHC）. The total content of m⁶A in hippocampus was determined by high performance liquid chromatography tandem mass spectrometry （LC-MS/MS）. ResultCompared with the conditions in the blank group， the average latency duration in the model group was prolonged， and the number of entries in the target quadrant， the target quadrant duration and the number of crossing the platform were decreased （P<0.01）. Additionally， the model group had seriously damaged structure of hippocampal CA1 and CA3 neurons， swollen mitochondria， expanded endoplasmic reticulum， and small number of synapses with some having blurred structure， and the expression levels of METTL3， METTL14， FTO， ALKBH5 as well as the total m⁶A level were lower than those in the blank group （P<0.05，P<0.01）. Compared with the model group， the Cuscutae Semen group had shortened average latency duration， increased number of entries in the target quadrant， target quadrant duration and number of crossing the platform （P<0.01）， alleviated damage of hippocampal CA1 and CA3 neurons， fine structure of mitochondrial and endoplasmic reticulum， and clear， intact and dense synapses. And the expression levels of METTL3， METTL14， FTO as well as the total level of m⁶A were up regulated， while the expression level of ALKBH5 was down regulated in the Cuscutae Semen group （P<0.05，P<0.01）. ConclusionCuscutae Semen improved the learning and memory ability of the offspring of the rats affected by fear damaging kidney during pregnancy， protected hippocampal neurons， and up-regulated the expression levels of METTL3， METTL14， FTO and the total m⁶A level in hippocampus.

One major strategy to generate genetically modified mouse models is gene targeting in mouse embryonic stem (ES) cells, which is used to produce gene-targeted mice for wide applications in biomedicine. However, a major bottleneck in this approach is that the robustness of germline transmission of gene-targeted ES cells can be significantly reduced by their genetic and epigenetic instability after long-term culturing, which impairs the efficiency and robustness of mouse model generation. Recently, we have established a new type of pluripotent cells termed extended pluripotent stem (EPS) cells, which have superior developmental potency and robust germline competence compared to conventional mouse ES cells. In this study, we demonstrate that mouse EPS cells well maintain developmental potency and genetic stability after long-term passage. Based on gene targeting in mouse EPS cells, we established a new approach to directly and rapidly generate gene-targeted mouse models through tetraploid complementation, which could be accomplished in approximately 2 months. Importantly, using this approach, we successfully constructed mouse models in which the human interleukin 3 (IL3) or interleukin 6 (IL6) gene was knocked into its corresponding locus in the mouse genome. Our study demonstrates the feasibility of using mouse EPS cells to rapidly generate mouse models by gene targeting, which have great application potential in biomedical research.