Objective To analyze the epidemiological characteristics and spatial-temporal distribution of drug-resistant tuberculosis in Pudong New Area from 2016 to 2021 and to provide scientific basis for the prevention and control of drug-resistant tuberculosis. Methods The data of tuberculosis patients in Pudong New Area from 2016 to 2021 were collected through the Tuberculosis Information Management System of China Disease Prevention and Control Information System. The geographic information database was established by ArcGIS software and the vector map of Pudong New Area for trend surface analysis and spatial autocorrelation analysis . Results From 2016 to 2021, a total of 3916 patients with etiological positive tuberculosis were found to be drug resistant with the drug resistance rate of 13.13%. The drug resistance rate of each year showed a decreasing trend (χ²_trend=14.917, P<0.001). The rates of drug resistance of male, retiree, age 50~<60 years old, 60~<70 years old and recurrent patient were higher. From 2016 to 2021, the incidence of drug-resistant TB showed no spatial aggregation.In the south - north direction, the north is higher than the south generally. In the east - west direction, the west is higher than the east generally. Conclusion Drug resistance screening should be strengthened for men, retirees, over 50 years old, and recurrent TB patients,. Prevention and control measures should be strengthened in street towns with dense population and large floating population.

Background::The goal of the assisted reproductive treatment is to transfer one euploid blastocyst and to help infertile women giving birth one healthy neonate. Some algorithms have been used to assess the ploidy status of embryos derived from couples with normal chromosome, who subjected to preimplantation genetic testing for aneuploidy (PGT-A) treatment. However, it is currently unknown whether artificial intelligence model can be used to assess the euploidy status of blastocyst derived from populations with chromosomal rearrangement.Methods::From February 2020 to May 2021, we collected the whole raw time-lapse videos at multiple focal planes from in vitro cultured embryos, the clinical information of couples, and the comprehensive chromosome screening results of those blastocysts that had received PGT treatment. Initially, we developed a novel deep learning model called the Attentive Multi-Focus Selection Network (AMSNet) to analyze time-lapse videos in real time and predict blastocyst formation. Building upon AMSNet, we integrated additional clinically predictive variables and created a second deep learning model, the Attentive Multi-Focus Video and Clinical Information Fusion Network (AMCFNet), to assess the euploidy status of embryos. The efficacy of the AMCFNet was further tested in embryos with parental chromosomal rearrangements. The receiver operating characteristic curve (ROC) was used to evaluate the superiority of the model. Results::A total of 4112 embryos with complete time-lapse videos were enrolled for the blastocyst formation prediction task, and 1422 qualified blastocysts received PGT-A ( n = 589) or PGT for chromosomal structural rearrangement (PGT-SR, n = 833) were enrolled for the euploidy assessment task in this study. The AMSNet model using seven focal raw time-lapse videos has the best real-time accuracy. The real-time accuracy for AMSNet to predict blastocyst formation reached above 70% on the day 2 of embryo culture, and then increased to 80% on the day 4 of embryo culture. Combing with 4 clinical features of couples, the AUC of AMCFNet with 7 focal points increased to 0.729 in blastocysts derived from couples with chromosomal rearrangement. Conclusion::Integrating seven focal raw time-lapse images of embryos and parental clinical information, AMCFNet model have the capability of assessing euploidy status in blastocysts derived from couples with chromosomal rearrangement.

Objective:To investigate the effect and mechanism of type ⅩⅦ collagen (COL17) on hair growth in mice with androgenetic alopecia (AGA).Methods:Forty-eight C57BL/6J mice were used to establish AGA model (the back hair of the mice was removed and dihydrotestosterone solution was applied) and divided into 6 groups of 8 mice each by random number table. Negative control group, injection of saline in the depilated area (single point injection of 0.05 ml, 5 points in total); positive control group, topical application of 5% minoxidil tincture in the depilated area, 1 ml/d; COL17 low, medium and high concentration groups, injection of 0.5, 1.0 and 2.0 mg/ml COL17 in the depilated area respectively (single point injection of 0.05 ml, 5 points in total); type Ⅲ and ⅩⅦ collagen (COL3+ COL17) combined high concentration group, injection of 2.0 mg/ml COL3 and COL17 in the depilated area (single point injection of 0.05 ml, 5 points in total). The total treatment time was 21 days, during which the hair growth of mice in each group was observed and recorded. After 21 days, the skin and subcutaneous tissue in the depilated area of the mice were taken to make pathological sections for HE staining, and the number and morphological changes of hair follicles were observed; fresh skin tissue in the depilated area of the mice was taken for total RNA sequencing analysis, and the differentially co-expressed genes were annotated by gene ontology (GO) functional annotation, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and gene set enrichment analysis (GSEA).Results:After 21 days of treatment, compared with the negative control group, the depilation area on the back of the mice in the positive control group, COL17 high concentration group, and COL3+ COL17 combined high concentration group was significantly reduced, and HE staining showed that the number of hair follicles was also significantly increased. Pearson correlation analysis, principal component analysis and cluster heat map between groups showed that COL17 high concentration group had high gene correlation with the positive control group ( R2=0.95, P=0.024), and the gene expression was relatively close, with 3 882 differentially expressed genes (1 705 up-regulated and 2 177 down-regulated) in the two groups, while COL3+ COL17 combined high concentration group had the highest gene correlation with the positive control group ( R2=0.96, P=0.001), and the gene expression was the closest, with 1 289 differentially expressed genes (385 up-regulated and 904 down-regulated). KEGG analysis showed that compared with the negative control group, the positive control group, COL17 high concentration group and COL3+ COL17 combined high concentration group of mice all upregulated Wnt signaling pathway, cell adhesion molecules and hedgehog signaling pathway related to hair growth. GO enrichment analysis suggested that COL17 high concentration group and COL3+ COL17 combined high concentration group had upregulated genes related to skin development and hair cycle. GSEA enrichment analysis found that COL17 high concentration group had upregulated genes related to fibroblast proliferation and interleukin-1 secretion, while COL3+ COL17 combined high concentration group had upregulated genes related to fibroblast migration, clearance of apoptotic cells and accelerated metabolism of reactive oxygen species. Conclusion:Local injection of 2.0 mg/ml COL17 has a certain promoting effect on hair growth in AGA model mice, and the effect is more significant after combined injection of 2.0 mg/ml COL3. Activation of Wnt signaling pathway is one of the main mechanisms of COL17 promoting hair growth.

Objective:To investigate the effect and mechanism of type ⅩⅦ collagen (COL17) on hair growth in mice with androgenetic alopecia (AGA).Methods:Forty-eight C57BL/6J mice were used to establish AGA model (the back hair of the mice was removed and dihydrotestosterone solution was applied) and divided into 6 groups of 8 mice each by random number table. Negative control group, injection of saline in the depilated area (single point injection of 0.05 ml, 5 points in total); positive control group, topical application of 5% minoxidil tincture in the depilated area, 1 ml/d; COL17 low, medium and high concentration groups, injection of 0.5, 1.0 and 2.0 mg/ml COL17 in the depilated area respectively (single point injection of 0.05 ml, 5 points in total); type Ⅲ and ⅩⅦ collagen (COL3+ COL17) combined high concentration group, injection of 2.0 mg/ml COL3 and COL17 in the depilated area (single point injection of 0.05 ml, 5 points in total). The total treatment time was 21 days, during which the hair growth of mice in each group was observed and recorded. After 21 days, the skin and subcutaneous tissue in the depilated area of the mice were taken to make pathological sections for HE staining, and the number and morphological changes of hair follicles were observed; fresh skin tissue in the depilated area of the mice was taken for total RNA sequencing analysis, and the differentially co-expressed genes were annotated by gene ontology (GO) functional annotation, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and gene set enrichment analysis (GSEA).Results:After 21 days of treatment, compared with the negative control group, the depilation area on the back of the mice in the positive control group, COL17 high concentration group, and COL3+ COL17 combined high concentration group was significantly reduced, and HE staining showed that the number of hair follicles was also significantly increased. Pearson correlation analysis, principal component analysis and cluster heat map between groups showed that COL17 high concentration group had high gene correlation with the positive control group ( R2=0.95, P=0.024), and the gene expression was relatively close, with 3 882 differentially expressed genes (1 705 up-regulated and 2 177 down-regulated) in the two groups, while COL3+ COL17 combined high concentration group had the highest gene correlation with the positive control group ( R2=0.96, P=0.001), and the gene expression was the closest, with 1 289 differentially expressed genes (385 up-regulated and 904 down-regulated). KEGG analysis showed that compared with the negative control group, the positive control group, COL17 high concentration group and COL3+ COL17 combined high concentration group of mice all upregulated Wnt signaling pathway, cell adhesion molecules and hedgehog signaling pathway related to hair growth. GO enrichment analysis suggested that COL17 high concentration group and COL3+ COL17 combined high concentration group had upregulated genes related to skin development and hair cycle. GSEA enrichment analysis found that COL17 high concentration group had upregulated genes related to fibroblast proliferation and interleukin-1 secretion, while COL3+ COL17 combined high concentration group had upregulated genes related to fibroblast migration, clearance of apoptotic cells and accelerated metabolism of reactive oxygen species. Conclusion:Local injection of 2.0 mg/ml COL17 has a certain promoting effect on hair growth in AGA model mice, and the effect is more significant after combined injection of 2.0 mg/ml COL3. Activation of Wnt signaling pathway is one of the main mechanisms of COL17 promoting hair growth.

Pleomorphic adenoma (PA) is the most common benign tumour in the salivary gland and has high morphological complexity. However, the origin and intratumoral heterogeneity of PA are largely unknown. Here, we constructed a comprehensive atlas of PA at single-cell resolution and showed that PA exhibited five tumour subpopulations, three recapitulating the epithelial states of the normal parotid gland, and two PA-specific epithelial cell (PASE) populations unique to tumours. Then, six subgroups of PASE cells were identified, which varied in epithelium, bone, immune, metabolism, stemness and cell cycle signatures. Moreover, we revealed that CD36⁺ myoepithelial cells were the tumour-initiating cells (TICs) in PA, and were dominated by the PI3K-AKT pathway. Targeting the PI3K-AKT pathway significantly inhibited CD36⁺ myoepithelial cell-derived tumour spheres and the growth of PA organoids. Our results provide new insights into the diversity and origin of PA, offering an important clinical implication for targeting the PI3K-AKT signalling pathway in PA treatment.
Humans ; Adenoma, Pleomorphic/genetics* ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; Transcriptome ; Myoepithelioma

Although somatic cells can be reprogrammed to pluripotent stem cells (PSCs) with pure chemicals, authentic pluripotency of chemically induced pluripotent stem cells (CiPSCs) has never been achieved through tetraploid complementation assay. Spontaneous reprogramming of spermatogonial stem cells (SSCs) was another non-transgenic way to obtain PSCs, but this process lacks mechanistic explanation. Here, we reconstructed the trajectory of mouse SSC reprogramming and developed a five-chemical combination, boosting the reprogramming efficiency by nearly 80- to 100-folds. More importantly, chemical induced germline-derived PSCs (5C-gPSCs), but not gPSCs and chemical induced pluripotent stem cells, had authentic pluripotency, as determined by tetraploid complementation. Mechanistically, SSCs traversed through an inverted pathway of in vivo germ cell development, exhibiting the expression signatures and DNA methylation dynamics from spermatogonia to primordial germ cells and further to epiblasts. Besides, SSC-specific imprinting control regions switched from biallelic methylated states to monoallelic methylated states by imprinting demethylation and then re-methylation on one of the two alleles in 5C-gPSCs, which was apparently distinct with the imprinting reprogramming in vivo as DNA methylation simultaneously occurred on both alleles. Our work sheds light on the unique regulatory network underpinning SSC reprogramming, providing insights to understand generic mechanisms for cell-fate decision and epigenetic-related disorders in regenerative medicine.
Male ; Mice ; Animals ; Cellular Reprogramming/genetics* ; Tetraploidy ; Pluripotent Stem Cells/metabolism* ; Induced Pluripotent Stem Cells/metabolism* ; DNA Methylation ; Spermatogonia/metabolism* ; Germ Cells/metabolism*

Nosiheptide is a typical thiopeptide antibiotic displaying potent activity toward various drug-resistant strains of Gram-positive pathogens.Although nosiheptide lacks in vivo activity, and good water-solubility with a series of uncontrollable analogues, which may limit its clinical application, glycosylated analogues may overcome problem of low activity and may improve its druggability.In search of novel glycosylated nosiheptide producers, we applied a genome mining strategy that identified Actinoalloteichus sp.AHMU CJ021 that contains all genes required.However, despite the presence of a predicted glycosyltransferase, glycosylated derivatives of nosiheptide were not detected, after following one strain many compounds (OSMAC) strategy and heterologous expression of a regulatory protein NocP.Nevertheless, nosiheptide produced by this strain was remarkably pure, and further experiments were conducted to improve its production by optimization of the culture medium.Under optimal conditions, 58.73 mg/L nosiheptide was produced, representing an almost 6-fold improvement compared to the original fermentation medium.Therefore, we consider Actinoalloteichus sp.AHMU CJ021 a suitable potential candidate for industrial production of nosiheptide, which provides the basis for solving the problem of nosiheptide structural analogues.

Objective:To investigate the clinical and imaging features of bilateral Wallerian degeneration of the middle cerebellar peduncles secondary to isolated pontine infarction.Methods:Patients diagnosed as bilateral Wallerian degeneration of cerebellar middle peduncle after isolated pontine infarction admitted to the Second Affiliated Hospital of Zhengzhou University from June 2017 to December 2021 were retrospectively included. Patients with bilateral Wallerian degeneration of cerebellar middle peduncle after isolated pontine infarction reported between January 2001 and December 2021 were collected by searching Chinese and English databases, and their clinical and imaging characteristics were summarized.Results:A total of 48 patients with bilateral Wallerian degeneration of cerebellar middle peduncle after isolated pontine infarction were included, including 14 patients admitted to the Second Affiliated Hospital of Zhengzhou University, and 34 patients collected by searching the Chinese and English databases. Thirty-three patients were males (68.75%) and 15 were females (31.25%). Their age was 65.8±10.7 years old (range, 37-88 years). Most patients had vascular risk factors, and hypertension was the most common. Dysarthria and limb weakness were the main clinical symptoms at admission. The infarct sites of all 48 patients were located in the blood supply area of paramedian pontine arteries, of which 37 (77.08%) were unilateral (18 on the left and 19 on the right), 6 (12.50%) were bilateral sides, and 5 (10.42%) had incomplete data. When Wallerian degeneration was diagnosed, 8 patients (16.67%) had dizziness or ataxia, 6 (12.50%) had aggravated original symptoms, and the remaining 34 (70.83%) had no new symptoms or aggravated original symptoms. All patients showed symmetrical abnormal signals in bilateral middle cerebellar peduncles, with obvious hyperintensity on T 2 or diffusion-weighted imaging (DWI). One patient showed T 2 hyperintensity in bilateral middle cerebellar peduncle on the next day after the onset of the infarction, which was the earliest case to find secondary Wallerian degeneration after isolated pontine infarction. Conclusions:Wallerian degeneration should be considered when symmetrical lesions of bilateral middle cerebellar peduncles occur after isolated pontine infarction. Wallerian degeneration may occur early after isolated pontine infarction. Most cases have no new symptoms or aggravated original symptoms. Conventional MRI can identify it early.

【Objective】 To analyze the polymorphisms of GYPA and GYPB mRNA spliceosomes associated with MNS blood group, and to explore the mechanism of subcellular localization of GPA and GPB protein isomerism encoded by various spliceosomes as well as the expression of MNS blood group antigen. 【Methods】 Ten blood samples of voluntary blood donors were randomly selected. The total mRNA of peripheral blood was extracted and reversed into cDNA. Nested PCR was used to amplify reading open frame of GYPA and GYPB gene, and sequencing was performed by Sanger. The base sequence obtained was compared with GYPA(NCBI: NM_002099) and GYPB(NCBI: Nm_002100.5). After the wild type and various splicing isomer of the open reading frame of GYPA and GYPB had been obtained, they were fused with the encoding gene of green fluorescent protein (GFP) by fusion PCR technology, then cloned and transfected into HEK293 cells for over expression. The subcellular localization of GPA-GFP and GPB-GFP fused fluorescent proteins was monitored by focusing laser scanning microscope. 【Results】 Exon-1 and Exon-2 were missing in GYPA mRNA of the 2 samples, and 2~26 amino acids were missing in the predicted GPA isomer, and the full length sequence of GYPB mRNA was complete. GYPA mRNA was intact in 6 samples, exon-2 was missing in GYPB mRNA, 13~45 amino acids were missing in the predicted GPB protein isomer, and other exon sequences were intact. One sample had intact GYPA mRNA, and 364~385 bases in exon-5 of GYPB mRNA were replaced by AG, indicating truncation of amino acid signal peptide. The GYP mRNA sequences of other samples were complete. The fluorescence signal of GP-GFP fusion protein showed that all GPA and GPB glycoprotein isomers, cloned according to various RNA splicing, could demonstrate the orientation distribution on the cell membrane surface, while some alternative splicing leaded to different degrees of protein dispersion in the cell, and affected the distribution speed and proportion of protein on the cell surface, which might be one of the reasons for the strength variation of MNS antigen. 【Conclusion】 The GYP mRNA spliceosome is obviously polymorphic, but the partial deletion of GYP mRNA fragment does not affect the localization and distribution of the protein isomers encoded by GYP mRNA on the cell surface, which can ensure the expression of MNS antigen characteristics.

Objective:To develop a comprehensive ability evaluation system for general practice management post in community health service institutions.Methods:Based on the previous studies and literature search on competency of general practitioners in various clinical posts, the first draft of the comprehensive ability indicator system for general management posts was designed. From November 2019 to January 2020, the comprehensive ability indicator system, classification and scoring method and evaluation method for general management posts were determined through three rounds of Delphi consultation.Results:Seventeen experts with administrative management experiences were invited for consultation, including 3 directors in charge and 14 general practice managers. The response rate of valid questionnaires in the three rounds of consultation were all 17/17, the authority of experts were all>0.7, and the coordination coefficients of expert opinions were 0.142, 0.212, and 0.532 (all P<0.01). The 11 evaluation indexes of the comprehensive ability of community general management posts were: achievement orientation, initiative, interpersonal understanding, service consciousness, influence, organization, training others, teamwork, team leadership, professional skills and confidence. Methods suitable for evaluation the ability of psychological test indicators were achievement orientation, interpersonal understanding, service consciousness and self-confident;appropriate indicators for no-leader group discussion were initiation, influence, team work, team leadership;appropriate indicators for in-tray test ability were organized cognition, professional skills;and the appropriate indicator for the ability of the structured interview was training others. Conclusions:The comprehensive ability evaluation system for community general management posts established in this study is scientific and practical, which may provide objective standard basis for the ability evaluation of community health management talents.