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OBJECTIVE: Bietti crystalline dystrophy (BCD) is a rare degenerative eye disease caused by mutations in the CYP4V2 gene, and Cyp4v3 is the murine ortholog to CYP4V2. To better understand the molecular pathogenesis of this disease and to explore the potential treatment we have established a Cyp4v3 knock-out mouse model. METHODS: Cyp4v3(-/-) mice were generated by clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 in embryonic stem cells of C57BL/6J mice. Ocular morphologic characteristics were evaluated via fundus imaging, histologic analysis of rods and cones via immunofluorescence, and phalloidin stain to observe retinal pigment epithelium (RPE) in whole-mounts, electroretinogram (ERG) was also conducted to examine the retinal function. RESULTS: The characteristic features of BCD recurred in the Cyp4v3(-/-) mice, including retinal crystalline deposits, atrophy and degeneration of RPE cells, and ERG amplitude decline of dark and light adapted a- and b- wave; however, the immunofluorescence stain of rod and cone cells did not show obvious differences when compared with the wild type (WT) mice. In the early stage of the disease, no crystal-like deposits were found in the fundus, ERG detection of the retinal function did not find a significant decline, and the morphological structure and quantity of the neural retina and RPE did not change significantly. Crystalline deposits occurred and converged when the Cyp4v3(-/-) mice at the end of 6 months, and the deposits disappeared when the Cyp4v3(-/-) mice at the end of 12 months. The ERG amplitude started to decline when the Cyp4v3(-/-) mice at the end of 6 months and deteriorated at the end of 12 months. The RPE cells of the 12-month old Cyp4v3(-/-) mice showed irregular shape by phalloidin staining of F-actin. The Cyp4v3(-/-) mice behaved normally and were viable and fertile when maintained under specific pathogen-free (SPF) housing conditions. CONCLUSION: Just like BCD patients, the disease progress of Cyp4v3(-/-) mouse is correlated with the age, which provides a good model for pathogenesis and gene therapy study in the future. The atrophy and degeneration of RPE take the lead in progressing of the disease, but the mechanism is not clear yet.

OBJECTIVE: To evaluate and compare whole exome sequencing (WES) and targeted panel sequencing in the clinical molecular diagnosis of the Chinese families affected with inherited retinal dystrophies (IRDs). METHODS: The clinical information of 182 probands affected with IRDs was collected, including their family history and the ophthalmic examination results. Blood samples of all probands and their relatives were collected and genomic DNA was extracted by standard protocols. The first 91 cases were subjected to the WES and the other 91 cases were subjected to a specific hereditary eye disease enrichment panel (HEDEP) designed by us. All likely pathogenic and pathogenic variants in the candidate genes were determined by Sanger sequencing and co-segregation analyses were performed in available family members. Copy number variations (CNVs) detected by HEDEP were further validated by multiplex ligation-dependent probe amplification (MLPA). As PRGR ORF15 was difficult to capture by next generation sequencing (NGS), all the samples were subjected to Sanger sequencing for this region. All sequence changes identified by NGS were classified according to the American College of Medical Gene-tics and Genomics and the Association for Molecular Pathology (ACMG/AMP) variant interpretation guidelines. In this study, only variants identified as pathogenic or likely pathogenic were included, while those variants of uncertain significance, likely benign or benign were not included. RESULTS: In 91 cases with WES, pathogenic or likely pathogenic variants were determined in 30 cases, obtaining a detection rate of 33.00% (30/91); While in 91 cases with HEDEP sequencing, pathogenic or likely pathogenic variants were determined in 51 cases, achieving the diagnostic rate of 56.04% (51/91), and totally, the diagnostic rate was 44.51%. HEDEP had better sequencing coverage and read depth than WES, therefore HEDEP had higher detection rate. In addition, HEDEP could detect CNVs. In this study, we detected disease-causing variants in 29 distinct IRD-associated genes, USH2A, ABCA4 and RPGR were the three most common disease-causing genes, and the frequency of these genes in Chinese IRDs population was 11.54% (21/182), 6.59% (12/182) and 3.85% (7/182), respectively. We found 43 novel variants and 6 cases carried variants in RPGR ORF15. CONCLUSION: NGS in conjunction with Sanger sequencing offers a reliable and effective approach for the genetic diagnosis of IRDs, and after evaluating the pros and cons of the two sequencing methods, we conclude that HEDEP should be used as a first-tier test for IRDs patients, WES can be used as a supplementary molecular diagnostic method due to its merit of detecting novel IRD-associated genes if HEDEP or other methods could not detect disease-causing va-riants in reported genes. In addition, our results enriched the mutational spectra of IRDs genes, and our methods paves the way of genetic counselling, family planning and up-coming gene-based therapies for these families.

本文分析了2例杓间区神经鞘瘤患者的临床表现及诊疗过程。例1患者声音嘶哑1年，咽喉部异物感，术前检查示杓间区占位，手术时先行支撑喉镜检查，术中冰冻怀疑为神经鞘瘤，再采用会厌根入路，切除杓间区肿瘤，修复创周黏膜，会厌复位缝合。例2患者声音嘶哑伴咽部不适，当地医院行支撑喉镜下喉肿物切除术并确诊为神经鞘瘤，术后2个月因复发转入我科，考虑既往手术史，直接采用会厌根入路切除，充分暴露、逐步分离并切除杓间区肿瘤，修复创周黏膜，缝合固定裂开的甲状软骨板。2例手术均顺利完成，术后随访患者发音良好，无复发。会厌根入路对该区域病变的手术是一种不错的选择。.

Triple-negative breast cancers (TNBC) are a subtype of breast cancer lacking of estrogen receptor (ER), progesterone receptor (PR), and human EGF-like receptor 2 (HER2). MiR-193 always acted as an oncogene and promoted toxic aldehyde accumulation and tyrosine hydroxylase dysfunction. The purpose of this study is to explore the function of miR-193 in triple-negative breast cancer.Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the mRNA level of miR-193 expression in 50 cases of TNBC tissues and para-cancerous specimens. Also, the relation between miR-193 level and the overall survival of TNBC patient was analyzed. MiR-193 mimic and miR-193 inhibitor oligos, as well as the corresponding negative control, were synthesized from RiboBio (Guangzhou, China).MiR-193 expression was higher in triple-negative breast cancer tissues and cell lines than the corresponding adjacent non-tumor tissues and normal cell lines. Upregulation of miR-193 predicted poor prognosis of TNBC patients. Overexpression of miR-193 promoted cell proliferation and invasion, while that was suppressed by the knockdown of miR-193. MiR-193 binds to the 3'-UTR of an inhibitor of growth family member 5 (ING5) mRNA to mediate the expression of ING5 in TNBC cells. The knockdown of miR-193 inhibited cell invasion-mediated epithelial-mesenchymal transition (EMT). Furthermore, the knockdown of miR-193 suppressed cell proliferation through the ING5/phosphatidylinositol 3-hydroxy kinase/protein kinase B (PI3K/AKT) signal pathway.MiR-193 enhanced cell invasion-mediated EMT and improved cell proliferation through the ING5/PI3K/AKT signal pathway in triple-negative breast cancer. The newly identified miR-193/ING5/PI3K/AKT axis provides novel insight into the pathogenesis of triple-negative breast cancer.

The study aimed to explore whether microRNA-155 and FOXP3 could regulate invasive and migratory capacities of colorectal cancer (CRC) cells by mediating Zinc finger E-box binding homeobox 2 (ZEB2) expression.Dual-luciferase reporter gene assay was performed to detect the binding condition between microRNA-155, FOXP3, and ZEB2. Protein and mRNA levels of ZEB2 in CRC cells were detected after overexpression of microRNA-155 and FOXP3 by Western blot and quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), respectively. In vitro experiments were conducted using HCT116 and SW620 cell lines. We first detected expression levels of microRNA-155, FOXP3, and ZEB2 in the normal colorectal epithelial cell line (NCM460) and CRC cell lines (HCT116 and SW620) by qRT-PCR. Protein expressions of ZEB2, E-cadherin, and vimentin in WT, LV-GFP, and LV-FOXP3 groups were detected. Wound healing assay and transwell assay were conducted to determine the regulatory effects of microRNA-155 and FOXP3 on invasive and migratory capacities of CRC cells, respectively.Dual-luciferase reporter gene assay found that FOXP3 binds to the promoter and intron regions of ZEB2, and microRNA-155 binds to the 3'UTR region of wild-type ZEB2. Overexpression of FOXP3 downregulated mRNA and protein levels of ZEB2. ZEB2 was highly expressed, whereas microRNA-155 and FOXP3 were lowly expressed in HCT116 and SW620 cells than NCM460 cells. MicroRNA-155 overexpression upregulated E-cadherin and downregulated vimentin in CRC cells. Overexpression of FOXP3 and microRNA-155 inhibited invasive and migratory capacities of CRC cells.MicroRNA-155 and FOXP3 can jointly regulate ZEB2 expression, thereby inhibiting the migration and invasion of colorectal cancer cells.

Chronic hepatitis B (CHB) is a major infectious disease which threatens people's health around the world. Health-related quality of life (HRQoL) has become an important index for evaluating the treatment and prognosis of patients with CHB. Many studies abroad have shown that the HRQoL of patients with CHB is influenced by various factors. Disease severity, clinical symptoms, and low self-efficacy are important influencing factors for patients' quality of life. Anti-viral therapy, proper physical therapy, or psychological intervention can significantly improve the HRQoL of community CHB patients, and HRQoL is even significantly improved in patients undergoing liver transplantation one year ago. These studies provide valid evidence for antiviral therapy for hepatitis B patients and liver transplantation techniques from the psychological level. At present, there still lacks studies on HRQoL of CHB patients in China. This article summarizes the measurement tools for HRQoL widely used in recent years in China and foreign countries, reviews the research advances in HRQoL in community patients with CHB, and discusses the perspectives for future studies.慢性乙型病毒性肝炎是危害全球人民生命健康的重大传染性疾病。健康相关生命质量（HRQoL）已成为评价慢性乙型肝炎患者治疗和预后的一个良好指标。国外许多研究结果表明慢性乙型肝炎患者的HRQoL受到不同程度的影响。病情的严重程度、临床症状和自我效能感不足等是影响患者生活质量的重要因素。对社区慢性乙型肝炎患者采取抗病毒治疗、适当的物理治疗或心理干预，都可以显著提高其HRQoL。甚至肝移植1年后的患者，HRQoL亦明显好转。这为乙型肝炎患者的抗病毒治疗和肝移植技术从心理学层面提供了有效证据。目前国内对慢性乙型肝炎患者的HRQoL研究尚处于空白阶段，现概述国内外近年广泛使用的HRQoL的测量工具，并对社区乙型肝炎患者的健康相关生命质量研究进展作一综述，以及对今后的相关研究进行讨论和展望。.

To investigate the common causes of death after drinking and the precautions for forensic identification.By reading alarm records, visiting investigations, on-site investigation transcripts, medical examination records and identification documents, the sex, age, corpse examination, toxic （drug） analysis and cause of death in 13 cases of post-drinking death were retrospectively analyzed.In 13 cases of post-drinking abnormal death, the causes of death were mainly drowning after drinking and asphyxia induced by stomach contents reflux. However, when accompanied by injury or other toxic poisoning, competition of death causes often occured.Combining the case and on-site inspection, a comprehensive systematic corpse examination and toxic （drug） analysis is of great significance for the forensic identification of cadavers of non-acute ethanol poisoning after drinking.酒后非正常死亡案例分析13例.探讨酒后死亡的常见死亡原因和法医学鉴定注意事项。.通过调阅报警记录、走访调查情况、现场勘验笔录、医学检验记录及鉴定文书等资料，对13例酒后死亡者的性别、年龄、尸体检验、毒（药）物检验和死亡原因等情况进行回顾性分析。.13例酒后非正常死亡案例中，死者均为男性，死亡原因以酒后溺水、胃内容物反流导致窒息多见，但在伴有损伤或其他毒物中毒时，常常导致死因竞争。.结合案情、现场勘验，进行全面系统的尸体检验和毒（药）物检验，对酒后非急性乙醇中毒死亡尸体的法医学鉴定有重要意义。.法医病理学；饮酒；死亡原因；案例分析.

In order to evaluate the effects of dietary taurine supplementation on visual fatigue induced by visual display terminals (VDT) work, 25 male college students aged from 20 to 24 years who were not engaged in VDT work were selected to participate in the study. Volunteers were randomly assigned to either the taurine supplementation (n=13) or the placebo supplementation control group (n=12). Before and after 12 days of taurine (3 g/day) or placebo supplementation, two identical 2.5-hr VDT work tests were performed while recording the P100, N75 and N145 latencies and P100 amplitude of pattern visual evoked potential (PVEP) and the frequency of critical flicker fusion (CFF). Following 2.5-hr of VDT work, the P100 and N75 latencies of PVEP increased ( P

To explore the radiosensitivity of andrographolide on esophageal cancer cell line ECA109. The inhibition effects of andrographolide were measured using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay. Clonogenic survival assay was used to evaluate the effects of andrographolide on the radiosensitivity of esophageal cancer cells. Immunofluorescence was employed to examine Bax expression. The changes in cell cycle distribution and apoptosis were assayed using flow cytometry. The expression of NF-κb/Cleaved-Caspase3/Bax/Bcl-2 was measured using Western blot analysis. DNA damage was detected via γ-H2AX foci counting. With a clear dose and time effects, andrographolide was found to inhibit the proliferation of esophageal cell line ECA109. The results of the clonogenic survival assay show that andrographolide could markedly enhance radiosensitivity (P0.05) with a sensitizing enhancement ratio of 1.28. Andrographolide caused a dose-dependent increase in Cleaved-Caspase3/Bax protein expression and a decrease in Bcl-2/NF-κb expression. Apoptosis in andrographolide-treated ECA-109 increased significantly compared with the apoptosis in the simple drug and radiation combined with drug groups (P0.001; P0.05). Moreover, compared with the independent radiation group, the andrographolide combined with radiation group increased the number of DNA double chain breaks. Andrographolide can increase the radiosensitivity of esophageal cell line ECA109. This result may be associated with the decrease in the NF-κb level and the induced apoptosis of esophageal cancer cells.

SummaryWhat is known and objective The glycopeptide antibiotics, vancomycin and teicoplanin, are the mainstay of therapy for severe gram-positive organisms such as methicillin-resistant Staphylococcus aureus. We report a case of Stevens–Johnson syndrome (SJS) induced by sequential therapy with teicoplanin and vancomycin, in a patient with chronic obstructive pulmonary disease (COPD). Case summary A 74-year-old Han Chinese with 1-year history of COPD was admitted for treatment of infective endocarditis. After teicoplanin therapy for 12 days, he developed pruritus and maculopapular over his trunk and limbs. His rash spread rapidly to most parts of the body surface area, 7 days after his anti-infection therapy was switched to vancomycin. This was stopped, but he developed SJS when teicoplanin was re-introduced. This patient recovered from his drug eruptions when both teicoplanin and vancomycin were stopped. Pharmacogenetic analyses revealed he was heterozygous with respect to two variants (rs2844682 of MUC21 and rs750332 of BAG6). What is new and conclusion Cross-reactivity between vancomycin and teicoplanin is rare. SJS attributable to sequential treatment with these two antibiotics has not been reported previously. Care should be taken when prescribing vancomycin in patients with a previous documented skin eruption to teicoplanin, especially in those who carry any susceptibility alleles to SJS/TEN.

The CD31 antigen (PECAM-1) has been reported to be a stable marker for a human CD4 T-cell subpopulation unable to produce interleukin-4 (IL-4). We show here that CD31 expression is not stable inasmuch as CD4 T-cell lines and clones derived from cell-sorted neonatal CD31+ cells lose CD31 upon repetitive cycles of stimulation and IL-2 expansion. Moreover, various cytokines (IL-1 alpha, IL-4, IL-6, transforming growth factor-beta) fail to reinduce CD31 on CD31- clones. Whereas all CD31+ CD4 T cells rapidly express high levels of the CD45RO antigen and down-regulate the L-selectin antigen after priming, CD31 disappears more slowly because only part of the cells lose CD31 expression upon each cycle of stimulation. Loss of CD31 reflects a functional maturation of CD45RO+ cells since, in a system which favours the development of Th2 effectors, IL-4 is produced by CD31- but not CD31+ effector T cells, whereas interferon-gamma is produced by both types of cells. However, CD31 is not a Th1 marker since it is not expressed on several Th1 antigen-specific clones. We conclude that CD31 is a maturation marker expressed on the great majority of naive CD45RO- CD4 T cells and on a subset of CD45RO+ CD4 T cells that are at an intermediate stage of maturation.

It was recently demonstrated that naive human and mouse CD4 T cells release low but sufficient levels of interleukin (IL)-4 at priming to support their development into IL-4 producers. To determine whether this IL-4 is produced by a minor subset of cells, freshly isolated human naive CD4 T cells were directly cloned by limiting dilution in the absence of exogenous IL-4. More than 95% of neonatal and 60% of adult naive T cells seeded in single-cell cultures could be expanded upon stimulation with anti-CD3 mAb immobilized on CD32-B7.1 L cell transfectants in the presence of IL-2. All 171 clones derived from four neonates and two adults produced IL-4 and IL-5 at generally high levels. Like the allergen-specific human Th2 clones described in the literature, these T cell clones produced little or no interferon-gamma upon stimulation via their T cell receptor/CD3 complex, whereas they released high levels of this cytokine when activated with phorbol 12-myristate 13-acetate+ionomycin. Cells cloned and expanded in the presence of anti-IL4 + anti-IL-4R mAb produced much lower levels of IL-4 and IL-5. It is concluded that almost every single naive human CD4 T cell primed and expanded in the absence of exogenous IL-4 releases sufficient autocrine IL-4 to support its clonal expansion into high IL-4/IL-5 producers.

B lymphocytes from chronic lymphocytic leukaemia (B-CLL) patients express the two CD23 isoforms (type A and B), which differ only in their intracytoplasmic domain. The abnormal regulation of the CD23 antigen in response to IL-4, IFNs alpha and gamma results in CD23 over-expression on B-CLL cells. Our present study shows that the two CD23 isoforms are differentially and abnormally regulated on B-CLL cells. IL-4 selectively up-regulates CD23 type A mRNA in five different B-CLL patients, whereas in normal B cells it enhances CD23 type A and is the most potent inducer of type B. In contrast, phorbol esters (PMA) up-regulate both CD23 isoforms in the malignant B cells and specifically increases type B in normal B cells. We next postulated that cytokines other than IL-4 regulate CD23 B isoform in B-CLL cells and therefore examined the effect of IL-2, IFN-gamma and IFN-alpha. We found that the ability of a given cytokine to induce B-CLL growth (i.e. IL-2 and IFN alpha) is concurrent with a selective up-regulation of CD23 type B mRNA, whereas lymphokines that have no B cell growth activity (i.e. IL-4 and IFN-gamma) specifically increase CD23 type A mRNA. We next showed that IL-4 and IFN gamma prevent hydrocortisone-induced programmed cell death and that the rescued malignant B cells mainly express CD23 type A. Given that CD23 molecule has been reported to play a role in normal B cell proliferation and survival, it is therefore proposed that in B-CLL cells the expression of CD23 type A may be related to cell viability and that of type B to cell proliferation. These data suggest that the CD23 molecule may contribute to the physiopathology of the disease which is characterized by the accumulation of long-lived and slow-dividing monoclonal B cells.

Thirty-nine psoriasis (PS) cases were treated by using fumigation and inhalation of Huayin Powder, it's effect was observed and comparison with composite Qingdaiwan group 34 cases was made. The result, showed that the efficacy overall was 94.90% in the treatment group, and 61.80% in the control (P0.01); recurrence rate and side effects of treatment group were obviously lower than those in the control; the medium molecular substance (MMS) content of treated group was higher than that of control (P0.01); and significantly lower in the treated group before treatment in comparison with that after treatment (P0.05) as well as lower than that of control. Before treatment there was insignificant difference in circulatory immune complex(CIC) between the treatment and control group (P0.05); however, after Huayin Powder was given, the CIC of treated group was significantly lower than that of before, treatment (P0.01). It indicated that both Huayin powder and composite Qingdaiwan were effective in treating PS, but the former being obviously better than the latter.

The maturation of naive CD4 T cells into interleukin (IL)-4-producing effectors was shown to require the presence of IL-4 at priming, the cellular origin of which remains unclear. We demonstrate here that naive T cells themselves release IL-4 at very low levels that are nevertheless sufficient to promote their development into Th2-like cells. This conclusion is based on three observations: (1) highly purified human naive CD4 T cells, of neonatal or adult origin, develop into Th2 effectors upon repetitive cycles of stimulation with anti-CD3 monoclonal antibody (mAb) cross-linked to CD32-B7 transfected L fibroblasts followed by IL-2 expansion; (2) IL-4 protein is readily detectable in the concentrated supernatant fluids of priming cultures performed in the presence of anti-IL-4 receptor mAb; and (3) addition of anti-IL-4 or anti-IL-4 receptor mAb at priming markedly inhibits the acquisition of IL-4- and IL-5-producing capacity while enhancing that of interferon-gamma.

Our previous studies have indicated that naive human CD4+ T cells of neonatal or adult origin may be the initial source of IL–4 which is required for their development into Th2 effectors. In addition to minute amounts of IL–4, anti–CD3/B7.1–activated naive cells also release readily detectable levels of IL–13 and IFN–γ. The production of IL–4 and IL–13 by naive T cells is differentially regulated by TGF–β and IL–12. Shortly after activation, naive T cells express surface OX40, a TNF–R family member whose ligand (OX40L) is constitutively expressed on a subset of dendritic cells. Engagement of OX40 on activated naive T cells increases their expression of IL–4 and IL–13, suppresses that of IFN–γ and promotes their development into Th2–like effectors.