Objective: To study the clinicopathologic features, immunophenotype, characteristic FISH pattern and prognosis of renal cell carcinoma (RCC) associated with chromosome X inversion harboring gene fusions involving TFE3. Methods: Ten cases of NONO-TFE3 RCC and four cases of RBM10-TFE3 RCC were investigated at Nanjing Jinling Hospital from 2009 to 2016 by clinicopathological findings, immunohistochemistry, and genetic analysis. Results: Morphologically, the distinct pattern of secretory endometrioid subnuclear vacuolization was overlapped with clear cell papillary RCC, and often accompanied by sheets of epithelial cells in NONO-TFE3 RCC. Most cases of RBM10-TFE3 RCC presented with the biphasic feature that acinar, tubular and papillary patterns of epithelioid cells combined with sheets of small cells with "pseudorosette-like" architectures. In addition, cytoplasmic vacuolization, nuclear groove, and psammoma bodies were also observed. Immunohistochemically, all NONO-TFE3 RCC cases were immunoreactive for TFE3, CD10, RCC markers, and PAX8, and negative for CK7, Cathepsin K, Melan A, HMB45, Ksp-cadherin, vimentin, and CD117. All 4 cases of RBM10-TFE3 RCC showed moderate to strong immunoreactivity for TFE3, Cathepsin K, CD10, Ksp-cadherin, E-cadherin, P504s, RCC marker, PAX8, and vimentin but negative for TFEB, HMB45 and CK7. CKpan and Melan A were at least focally expressed. The antibody to Ki-67 showed labeling of 3%-8% (mean 5%). There were some expression discrepancies of immunochemistry between different histological patterns. PAX8, CKpan, P504s, and Ksp-cadherin were expressed in epithelioid areas but not in small-cell areas. Ki-67 labeling index of epithelioid areas was higher than that in small-cell areas. In molecular analysis, NONO-TFE3 fusion transcripts were identified in 6 patients. The fusion points were between exon 7 of NONO and exon 6 of TFE3 in 5 patients and between exon 9 of NONO and exon 5 of TFE3 in one patient. All 4 cases of RBM10-TFE3 RCC demonstrated to have RBM10-TFE3 fusion transcripts and the fusion points were between exon 5 of TFE3 and exon 17 of RBM10. Using TFE3 break-apart FISH assay, all 10 cases of NONO-TFE3 RCC showed characteristic patterns of equivocal split signals with a distance of nearly 2 signal diameters. All 4 cases of RBM10-TFE3 RCC showed colocalized or subtle split signals with a distance of <1 signal diameter, which was considered as negative results. Long-term follow-up was available for 7 patients of NONO-TFE3 RCC and 4 patients of RBM10-TFE3 RCC. All patients were alive with no evidence of disease. Conclusions Two rare genotypes, NONO-TFE3 RCC and RBM10-TFE3 RCC, are reported in this study. Both of these two tumors show specific morphology and good prognosis, along with the positive TFE3 staining and the equivocal or false-negative TFE3 FISH results, which could be missed. PCR detection or next-generation sequencing can determine the genotype.

Objective:To understand the genome-wide sequence variation and molecular evolution of coxsackievirus A4 (CV-A4) strain in Anhui province, so as to provide a theoretical basis for the pathogenic monitoring and scientific prevention and treatment of hand, foot and mouth disease in the future.Methods:Five CVA4 isolates of 2020 were sequenced by first-generation sequencing method. MEGA11.0 was used to construct a phylogenetic tree based on VP1 region for 5 CV-A4 isolates, 32 CV-A4 strains and Enterovirus A71(EV-A71) prototype strain BrCr, and the isolates and enterovirus A (EV-A) prototype strains based on P1, P2 and P3 regions respectively, and DNAStar was used for amino acid sequence comparison in VP1 region. BioEdit7.2 was used for amino acid displacement entropy analysis and amino acid site entropy mapping. SimPlot3.5 and RDP4 were used for recombination analysis of CV-A4 isolate and EV-A prototype representative strains, and DnaSP6 software was used for selection pressure analysis of isolates and reference representative strains.Results:The phylogenetic tree showed that the isolates belonged to the C2 subtype, which belonged to the same clade as the CV-A4 strain circulating in Chinese mainland, and the amino acid sequence homology of the C2 subtype branch was 97.3%-100%, and the isolates had 6 amino acid variation sites compared with the prototype. The selection pressure analysis showed that the CV-A4 strain of the C2 subtype was affected by negative selection pressure, and there were 25 mutagenic sites in the amino acid sequence in the coding region of the displacement entropy analysis, accounting for 1.14%, and the evolution of the strain mainly depended on recombination. Recombination analysis showed that the isolates recombined with a variety of EV-A prototype strains in the P2 and P3 regions, and the recombination section with the CV-A5 prototype strain was longer, especially in the 3A-3C section, and P1 was a relatively conserved region.Conclusions:CV-A4 has frequent recombination events with other EV-A prototype strains in P2 and P3, and the molecular evolution of CV-A4 in Anhui should continue to be closely monitored.

Hearing loss is one of the most common neurosensory disorders in humans,severely affecting pa-tients'quality of life with lack of ideal treatments.Its pathogenesis is related to oxidative stress,inflammation and apoptosis in the inner ear.Recent studies have demonstrated that members of the signal transducer and activator of transcriptions(STATs)family are involved in regulating gene expression in auditory cells during inner ear develop-ment and physiological activities such as apoptosis,oxidative stress,inflammation and autophagy during auditory disorders.In this paper,we review the research on STATs in inner ear development and hearing loss,and elucidate their specific molecular mechanisms,aiming to provide theoretical guidance and direction for the prevention and treatment of hearing loss.

Objective:To analyze the salary level, change trend and salary satisfaction of nurses in tertiary public hospitals in China.Methods:The data was derived from three round third-party evaluation of the China Healthcare Improvement Initiative at 136 tertiary public hospitals from December 2017 to January 2018, March 2019 and January to March 2021. The self-reported salary, expected salary and salary satisfaction rate of nurses were analyzed.Results:The sample sizes of the three evaluations were 27 575, 27 568 and 25 197, representing the salary situation in 2017, 2018 and 2020 respectively. In 2017, 2018 and 2020, the actual average annual salary of nurses was 81 600 yuan, 100 000 yuan and 110 000 yuan respectively, the expected average annual salary was 140 000 yuan, 160 000 yuan and 160 000 yuan respectively, and the ratio of expected salary to actual salary was 1.72, 1.60 and 1.45 respectively. In 2017, 2018 and 2020, the proportion of nurses satisfied with their current income was 34.0%, 33.7% and 43.6% respectively. In 2020, nurses in the eastern region, other specialized hospitals, senior professional titles and graduate degrees had highest annual salary.Conclusions:The salary level and satisfaction rate of nurses in tertiary public hospitals in China show a slow growth trend, and the gap between actual salary and expected salary is gradually narrowing.

Due to multiple factors such as the closed research and development (R&D) system and differences in international mainstream regulatory standards, the internationalization of Chinese patent medicines has been weakly promoted. Multi-case analysis based on grounded theory can identify the key factors that promote the internationalization of Chinese patent medicines in each link, so as to put forward suggestions for promoting the internationalization of Chinese patent medicines. By retrieving the research literature on the internationalization of Chinese patent medicines included in CNKI, and further searching the official websites of the corresponding enterprises, as well as the news reports, comments and analysis of related events on official websites such as Xinhua News Agency, People's Network and China Medical Device Network, 27 textual materials were obtained, 6 classic cases of international registration of Chinese patent medicines were summarized, and three-level coding was used to organize and analyze them level by level. 25 initial categories and 7 main categories were extracted, and 4 core categories were finally summarized: promote R&D and promotion in advantageous areas based on international needs, build a quality control system in line with the international standards, select appropriate national markets based on international demand, and carry out international collaborative R&D in the whole life cycle. Based on this, suggestions were put forward: in order to promote the internationalization process of Chinese patent medicines, priority disease types and product formulations should be determined based on demand during the drug discovery phase; full cycle international cooperation in drug R&D should be carried out; a Chinese patent medicines and simple preparations supervision system that is in line with the international standards in the link of drug quality supervision should be constructed; countries with flexible and experienced regulatory rules in the drug approval and marketing process should be chosen.

The pathological processes of Alzheimer's disease and type 2 diabetes mellitus have been demonstrated to be linked together. Both PDE9 inhibitors and PPAR agonists such as rosiglitazone exhibited remarkable preclinical and clinical treatment effects for these two diseases. In this study, a series of PDE9 inhibitors combining the pharmacophore of rosiglitazone were discovered. All the compounds possessed remarkable affinities towards PDE9 and four of them have the IC values <5 nmol/L. In addition, these four compounds showed low cell toxicity in human SH-SY5Y neuroblastoma cells. Compound , the most effective one, gave the IC of 1.1 nmol/L towards PDE9, which is significantly better than the reference compounds PF-04447943 and BAY 73-6691. The analysis of putative binding patterns and binding free energy of the designed compounds with PDE9 may explain the structure-activity relationships and provide evidence for further structural modifications.

Our previous study demonstrated that phosphodiesterase 8 (PDE8) could work as a potential target for vascular dementia (VaD) using a chemical probe 3a. However, compound 3a is a chiral compound which was obtained by chiral resolution on HPLC, restricting its usage in clinic. Herein, a series of non-chiral 9-benzyl-2-chloro-adenine derivatives were discovered as novel PDE8 inhibitors. Lead 15 exhibited potent inhibitory activity against PDE8A (IC₅₀ = 11 nmol/L), high selectivity over other PDEs, and remarkable drug-like properties (worthy to mention is that its bioavailability was up to 100%). Oral administration of 15 significantly improved the cAMP level of the right brain and exhibited dose-dependent effects on cognitive improvement in a VaD mouse model. Notably, the X-ray crystal structure of the PDE8A-15 complex showed that the potent affinity and high selectivity of 15 might come from the distinctive interactions with H-pocket including T-shaped π-π interactions with Phe785 as well as a unique H-bond network, which have never been observed in other PDE-inhibitor complex before, providing new strategies for the further rational design of novel selective inhibitors against PDE8.

Optimization efforts were devoted to discover novel PDE10A inhibitors in order to improve solubility and pharmacokinetics properties for a long-term therapy against pulmonary arterial hypertension (PAH) starting from the previously synthesized inhibitor

Vascular dementia (VaD) is the second commonest type of dementia which lacks of efficient treatments currently. Neuroinflammation as a prominent pathological feature of VaD, is highly involved in the development of VaD. In order to verify the therapeutic potential of PDE1 inhibitors against VaD, the anti-neuroinflammation, memory and cognitive improvement were evaluated in vitro and in vivo by a potent and selective PDE1 inhibitor 4a. Also, the mechanism of 4a in ameliorating neuroinflammation and VaD was systematically explored. Furthermore, to optimize the drug-like properties of 4a, especially for metabolic stability, 15 derivatives were designed and synthesized. As a result, candidate 5f, with a potent IC₅₀ value of 4.5 nmol/L against PDE1C, high selectivity over PDEs, and remarkable metabolic stability, efficiently ameliorated neuron degeneration, cognition and memory impairment in VaD mice model by suppressing NF-κB transcription regulation and activating cAMP/CREB axis. These results further identified PDE1 inhibition could serve as a new therapeutic strategy for treatment of VaD.

Objective: To study the effects of Phellodendron amurense on herpes simplex virus 1 (HSV-1) and cytokines, and to explore the mechanism of Phellodendron amurense inhibiting HSV-1 virus through multiple channels. Methods: Viruses were inoculated into medicine treated HeLa cells. The proliferation of virus was observed by fluorescence microscopy. The transcriptional levels of glycoprotein gD and functional protein US1 on the surface of virus envelope were detected by quantitative (q) PCR. After incubating HeLa cells for 24 h, qPCR was used to detect the expression of intrinsic immune factors such as IP-10, IL-12, IFN-gamma and transcription factor NF-kappa B (P65). The expression and nuclear location of NF-kappa B (P65) protein were detected by immunofluorescence. Results: Fluorescence showed that the proliferation of virus decreased significantly at 8 and 40 ng/ml (P<0.01), and the transcription levels of viral protein gD and US1 decreased (P<0.05). After incubation for 24 hours, the transcription levels of IP-10, IL-12 and IFN-gamma in HeLa cells increased significantly (P<0.01). The transcription level of transcription factor NF-kappa B (P65) also increased (P<0.05), and immunofluorescence showed that the nuclear penetration rate of p65 subunit of NF-kappa B (P65) in each group increased (P<0.05). Conclusions Phellodendron amurense extract can inhibit HSV-1 by inhibiting the transcription of viral functional protein and promoting the expression of cellular immune factors.