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Neutrophils are the most abundant circulating leukocytes and the first line of defense against invading pathogens. They are key components of the innate immune system. Neutrophils also cause tissue damage in various autoimmune and inflammatory diseases and play an important role in cancer progression. Due to the complex relationship between various diseases and neutrophils, these cells have become potentially important targets for therapeutic interventions. Monitoring neutrophils in the tumor microenvironment is critical for tumor treatment and prognostic analysis but remains challenging. Molecular imaging technology has made great progress as a valuable tool for noninvasively visualizing biological events and establishing effective cancer diagnoses and treatment methods. Molecular probes designed based on the characteristics of neutrophils, such as their flexible morphology, the abundance of surface receptors, and the absence of immunogenicity, have shown great potential. This has created an opportunity for novel ideas and research methods for the diagnosis and targeted therapy of inflammatory diseases and tumors, with the goal of integrated diagnosis and treatment. This review discusses the diverse tumor detection and diagnostic imaging strategies based on neutrophils. It is anticipated that neutrophil-based imaging will soon be gradually integrated into clinical applications.Neutrophils play a role in promoting/inhibiting tumor growth in the tumor microenvironment.To be more conducive to discovering neutrophils in tumors and guiding treatment, medical imaging technology has developed stepwise.In the context of the intersection of imaging technology, great breakthroughs have been made in the individualized research of tumor diagnosis.This article contributes to the promotion of new multimodal molecular imaging technology and the application of rapid diagnosis individualization.

BACKGROUND: Giant cell tumor of bone (GC), osteosarcoma (OS) and Ewing’s sarcoma (ES) are three different types of bone cancer with common and specific pathology features. OBJECTIVE: The purpose of the study was to examine the relationship and differences of the three bone tumors using clinical samples. METHODS: Through screening the profiles of clinical samples from GC, OS and ES patients using a humanoncology array, we found 26, 25 and 15 tumorigenesis factors significantly increased in GS, OS and ES tissues compared to normal individuals. eNOS, endostatin, HIF-1α, IL-6, CCL2/MCP-1, CCL8/MCP-2, CCL7/MCP-3, Tie and VEGF directly or indirectly involve in the metastasis Therefore, expression levels of the 6 factors were further determined by Western blot. RESULTS: The results showed levels of MCP1, MCP2, MCP3 or IL-6 in the GS, OS and ES significantly increased, and the expression levels of angiogenesis and anti-angiogenesis factors containing eNOS, endostatin, HIF-1α, Tie or VEGF were enhanced. CONCLUSIONS: Our results suggest that eNOS, endostatin, HIF-1α, IL-6, CCL2/MCP-1, CCL8/MCP-2, CCL7/MCP-3, Tie and VEGF may play important roles in tumorigenesis, reveal the expression differences of tumor-associated cytokines and angiogenesis related factors, and provide clinical evidence for studying the mechanisms on the metastasis in GC, OS and ES.

Circulating tumor DNA (ctDNA) correlates with tumor burden and provides early detection of treatment response and tumor genetic alterations in breast cancer. Neoadjuvant chemotherapy (NACT) has become standard therapy for local advanced breast cancer (LABC). The aim of our study was to investigate plasma ctDNA as a prognostic marker for outcome in patients with LABC treated with NACT. A total of 56 patients with LABC were involved in this study. ctDNA mutations were investigated by using a 100 gene panel-target capture next-generation sequencing. The patients then received standard NACT therapy: adriamycin and cyclophosphamide and paclitaxel (AC-T) or AC-TH (AC-T+ Trastuzumab) regimen. The efficacy of NACT was evaluated by Miller-Payne grading system. A predictive and weight model was used to screen ctDNA point mutation biomarkers for NACT. The ctDNA mutational profile of LABC patients was identified. For nonsynonymous mutations, the top 5 mutated genes were MTHFR (51/56, 91.1%), XPC (50/56, 89.3%), ABCB1 (48/51, 94.1%), BRCA2 (38/56, 67.9%), and XRCC1 (38/56, 67.9%). In addition, the mutation frequencies of PIK3CA and TP53 were 32.1% (18/56) and 26.8% (15/56), respectively. The predictive model indicated that XRCC1 44055726 (TG-) mutation (25/56, 44.6%) was significantly associated with Miller-Payne 4-5 and Miller-Payne 3-5 responses. While mTOR 11249132(GC) mutation (23/56, 41.1%) was associated with Miller-Payne 1-4 or Miller-Payne 1-3 responses. Furthermore, XRCC1 44055726 (TG-) accompanied by mTOR wild type predicted a good NACT efficacy in all response classification systems. The ROC curves to discriminate good neoadjuvant chemotherapy efficiency (Miller-Payne 4-5) and poor efficiency (Miller-Payne 1-3) were created, and AUC value was 0.77. Our results suggested that ctDNA mutation of XRCC1 44055726 (TG-) might be a positive biomarker for NACT therapy in LABC, while mTOR 11249132(GC) mutation was potentially associated with NACT resistance.

Mps one binder 2 (MOB2) regulates the NDR kinase family, however, whether and how it is implicated in cancer remain unknown. Here we show that MOB2 functions as a tumor suppressor in glioblastoma (GBM). Analysis of MOB2 expression in glioma patient specimens and bioinformatic analyses of public datasets revealed that MOB2 was downregulated at both mRNA and protein levels in GBM. Ectopic MOB2 expression suppressed, while depletion of MOB2 enhanced, the malignant phenotypes of GBM cells, such as clonogenic growth, anoikis resistance, and formation of focal adhesions, migration, and invasion. Moreover, depletion of MOB2 increased, while overexpression of MOB2 decreased, GBM cell metastasis in a chick chorioallantoic membrane model. Overexpression of MOB2-mediated antitumor effects were further confirmed in mouse xenograft models. Mechanistically, MOB2 negatively regulated the FAK/Akt pathway involving integrin. Notably, MOB2 interacted with and promoted PKA signaling in a cAMP-dependent manner. Furthermore, the cAMP activator Forskolin increased, while the PKA inhibitor H89 decreased, MOB2 expression in GBM cells. Functionally, MOB2 contributed to the cAMP/PKA signaling-regulated inactivation of FAK/Akt pathway and inhibition of GBM cell migration and invasion. Collectively, these findings suggest a role of MOB2 as a tumor suppressor in GBM via regulation of FAK/Akt signaling. Additionally, we uncover MOB2 as a novel regulator in cAMP/PKA signaling. Given that small compounds targeting FAK and cAMP pathway have been tested in clinical trials, we suggest that interference with MOB2 expression and function may support a theoretical and therapeutic basis for applications of these compounds.

Research question Can single-nucleotide polymorphisms (SNP) of genes related to progesterone synthesis predict the risk of premature serum progesterone elevation in women undergoing gonadotrophin-releasing hormone agonist protocols for ovarian stimulation? Design A total of 765 women were divided into high progesterone and normal progesterone groups according to progesterone concentration on the day of human chorionic gonadotrophin (HCG) administration, with the 75th percentile as the threshold between the group. Associations were analysed of genetic information from whole exome sequencing and the clinical characteristics of the two groups to identify the relationship between SNP, haplotypes and serum progesterone elevation. Results Among 40 common SNP of eight genes (FSHR, LHCGR, ESR1, ESR2, PGR, HSD3B1, CYP11A1 and CYP17A1), no statistical significance between the high and normal progesterone groups was identified in the distribution of genotypes and allele frequencies after multiple test correction to adjust the false discovery rate (PFDR > 0.05). When compared with the most common haplotypes of each gene, haplotype GAAG in CYP17A1 was associated with a 1.44-fold increased risk of progesterone elevation (95% confidence interval [CI] 1.22–1.69, PFDR Conclusions Polymorphism in genes involved in enzymes or hormone receptors in the progesterone synthesis pathway may have a role in modifying risk of serum progesterone elevation.

PURPOSE: To evaluate the efficacy of preimplantation genetic testing (PGT) for α- and β-double thalassemia combined with aneuploidy screening using next-generation sequencing (NGS). METHODS: An NGS-based PGT protocol was performed between 2017 and 2018 for twelve couples, each of which carried both α- and β-thalassemia mutations. Trophectoderm biopsy samples underwent whole-genome amplification using multiple displacement amplification (MDA), followed by NGS for thalassemia detection and aneuploidy screening. A selection of several informative single nucleotide polymorphisms (SNPs) established haplotypes. Aneuploidy screening was performed only on unaffected noncarriers and carriers. Unaffected and euploid embryos were transferred into the uterus through frozen-thawed embryo transfer (FET). RESULTS: A total of 280 oocytes were retrieved following 18 ovum pick-up (OPU) cycles, with 182 normally fertilized and 112 cultured to become blastocysts. One hundred and seven (95.5%, 107/112) blastocysts received conclusive PGT results, showing 56 (52.3%, 56/107) were unaffected. Thirty-seven (66.1%, 37/56) of the unaffected were also identified as euploid. One family had no transferable embryos. Unaffected and euploid embryos were then transferred into the uterus of the other 11 couples resulting in 11 healthy live births. The clinical pregnancy rate was 61.1% (11/18) per OPU and 68.8% (11/16) per FET, with no miscarriage reported. Seven families accepted the prenatal diagnosis and received consistent results with the NGS-based PGT. CONCLUSION: This study indicated that NGS could realize the simultaneous PGT of double thalassemia and aneuploidy screening in a reliable and accurate manner. Moreover, it eliminated the need for multiple biopsies, alleviating the potential damages to the pre-implanted blastocysts. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10815-020-01732-7) contains supplementary material, which is available to authorized users.

Glioma is the most common malignant tumor of the central nervous system, and it is characterized by high relapse and fatality rates and poor prognosis. Bufalin is one of the main ingredients of Chan-su, a traditional Chinese medicine (TCM) extracted from toad venom. Previous studies revealed that bufalin exerted inhibitory effects on a variety of tumor cells. To demonstrate the inhibitory effect of bufalin on glioma cells and glioma stem-like cells (GSCs) and discuss the underlying mechanism, the proliferation of glioma cells was detected by MTT and colony formation assays following treatment with bufalin. In addition, we investigated whether bufalin inhibits or kills GSCs using flow cytometry, Western blotting, and reverse transcription polymerase chain reaction analysis (RT-PCR). Finally, we investigated whether bufalin could improve the therapeutic effect of temozolomide (TMZ) and discussed the underlying mechanism. Taken together, our data demonstrated that bufalin inhibits glioma cell growth and proliferation, inhibits GSC proliferation, and kills GSCs. Bufalin was found to induce the apoptosis of GSCs by upregulating the expression of the apoptotic proteins cleaved caspase 3 and poly(ADP-ribose) polymerase (PARP) and by downregulating the expression of human telomerase reverse transcriptase, which is a marker of telomerase activity. Bufalin also improved the inhibitory effect of TMZ on GSCs by activating the mitochondrial apoptotic pathway. These results suggest that bufalin damages GSCs, induces apoptosis, and enhances the sensitivity of GSCs to TMZ.

Humans are at the core of the social-technical system, and their behavioral errors affect the reliability and safety of the entire system in varying degrees. Occupational accidents and large-scale industrial accidents are often attributed to human errors, accounting for more than 80% of accidents. In view of the complexity of systems and the coupling of elements, a new HFA method is proposed based on a complex network. According to system safety theory, a complex network is regarded as a network composed of humans, matters, environments, and management, and the basic structure of the HFA network is summarized. On this basis, a system safety method of HFA is developed which proposes a universal human error causation model. Moreover, a network analysis method for human errors is also presented, which is a comprehensive analysis of human errors that have occurred. Finally, the above methods are applied to gas explosion accidents that occurred in China. Results show that the two methods proposed are universal to all fields, and their combination improves the effectiveness of human error management and promotes the targeted, proactive, systematic, and dynamic prevention of critical nodes and paths from a holistic perspective.

Bone marrow mesenchymal stem cells (BMMSCs) are characterized by their pluripotent differentiation and self-renewal capability and have been widely applied in regenerative medicine, gene therapy, and tissue repair. However, inflammatory response after BMMSCs transplantation was found to impair the osteogenic differentiation of BMMSCs. Thus, understanding the mechanisms underlying inflammation response will benefit the clinical use of BMMSCs. In this study, using a cell model of TNF-α-induced inflammatory response, we found that TNF-α treatment greatly elevated intracellular oxidative stress and induced endoplasmic reticulum (ER) stress by elevating the expression levels of ER sensors, such as PERK, ATF6 and IRE1A. Oxidative stress and ER stress formed a feedback loop to mediate TNF-α-induced inflammation response in BMMSCs. Moreover, c-Jun N-terminal kinase (JNK) signal pathway that coupled to the ER stress was significantly activated by increasing its phosphorylation upon TNF-α treatment. Importantly, pharmacological inhibition of ER stress effectively eliminated the phosphorylation of JNK and attenuated the TNF-α-induced inflammation response. In conclusion, our results indicated that TNF-α induced oxidative and ER stress, thereby leading to JNK activation, and generating inflammation response in BMMSCs. This pathway underlying TNF-α-induced inflammation response may provide new strategies to improve BMMSCs osteogenesis and other inflammation-associated bone diseases.

To investigate the incidence of skin diseases, the factors affecting the life quality, anxiety, and depression of patients with skin diseases and the correlation among the 3. A total of 1127 patients with skin diseases were selected online from March 2018 to June 2018. Logistic regression was used to analyze the factors influencing life quality, anxiety, and depression. Spearman rank correlation was used to analyze the correction among the life quality, anxiety, and depression. There was no significant difference in ethnicity, education, and time of question among patients (P > .05). Sex ratio, age, marital status, and occupation were statistically significant difference among patients with different skin diseases (P

Insulin-like growth factor 1 (IGF-1), a multifunctional peptide that involves in cell proliferation and differentiation, can induce strong osteogenic differentiation in bone marrow mesenchymal stem cells (BMMSCs). However, it remains unknown whether intracellular Ca

Ubiquitin-specific protease 26 (USP26) encodes a predicted protein containing his- and cys- domains that are conserved among deubiquitinating enzymes. USP26 is specifically expressed in testis tissue and is a potential infertility gene. In the present study, we performed genetic testing related to spermatogenesis impairment in a patient with idiopathic severe oligozoospermia to identify the cause. The patient underwent clinical examination and reproductive hormone testing. Genes associated with male infertility, including USP26, were assessed by targeted exome sequencing. A novel frameshift mutation, c.2195delT (p.Phe732Serfs*14), was identified in USP26. This frameshift mutation was located in residue 732 of USP26 gene, leading to loss of the conserved deubiquitinating enzyme His-domain and producing a truncated protein of 744 amino acids. Bioinformatics analysis revealed this mutation to be pathogenic. A novel framshift mutation c.2195delT (p.Phe732Serfs*14) in USP26 gene was reported to be associated with male infertility in a Chinese patient with severe oligozoospermia.

Male infertility is a complex multifactorial disease affecting approximately 10% of couples who want to have children. Some cases of infertility can be explained by genetic factors. Septins are members of the GTPase superfamily, which are involved in diverse biological processes including morphogenesis, compartmentalization, cytokinesis, and apoptosis. The septin 12 gene, SEPT12, is expressed exclusively in post-meiotic male germ cells and is considered as a critical gene for spermatogenesis. In this study, we evaluated 200 patients with non-obstructive azoospermia and detected mutations of 25 spermatogenesis-associated genes by targeted exome sequencing. We report a missense SEPT12 variant, c.673C>A/p.Gln225Lys, in an infertile man with non-obstructive azoospermia. The variation was located inside the GTPase domain and had a SIFT score of 0.02 (

Cell-based immunotherapy holds promise in the quest for the treatment of cancer, having potential synergy with surgery, chemotherapy and radiotherapy. As a novel approach for adoptive cell-based immunotherapy, cytokine-induced killer (CIK) cells have moved from the 'bench to bedside'. CIK cells are a heterogeneous subset of ex-vitro expanded, polyclonal T-effector cells with both natural killer (NK) and T-cell properties, which present potent non-major histocompatibility complex-restricted cytotoxicity against a variety of tumor target cells. Initial clinical studies on CIK cell therapy have provided encouraging results and revealed synergistic antitumor effects when combined with standard therapeutic procedures. At the same time, issues such as inadequate quality control and quantity of CIK cells as well as exaggerated propaganda were continuously emerging. Thus, the Ministry of Health in China stopped CIK cell therapy in May 2016, which was a major setback for the innovation of CIK cell-based immunotherapy. Thus, it is very important to modify technical criteria to develop a standardized operation procedure (SOP) and standardized system for evaluating antitumor efficacy in a safe way.

MENK, an endogenous opioid peptide has been reported to have many immunological and antitumor activities. So far the detailed mechanisms of antitumor through regulating DCs by MENK have not been elucidated yet. The aim of this work was to investigate the antitumor mechanisms of MENK via regulating DC. The monitoring methods, such as ELISA, MTS assay, CFSE, Real-time PCR and Western blot were included in our research. We found bone marrow derived dendritic cells (BMDCs) in 36 female C57BL/6 mice treated with MENK enhanced expression of key surface molecules, increased production of critical cytokines reduced endocytosis of FITC-dextran, upregulated TLR4 through MyD88/NF-κB signaling pathway and mounted higher antitumor activity. These observations were further supported by an enhancement of nuclear translocation of the p65NF-κB subunit involved in this process. Surprisingly, mu-opioid receptors were the main participants of this kind of activation, not delta-opioid receptors nor kappa-opioid receptors, and these interactions could be partly blocked by Naltrexone (a kind of opioid antagonist). In vivo study the activated CD4 + , CD8 + T cells and decreased ability to induce differentiation of Foxp3 + regulatory T cells were detected post treatment of MENK. Thus, it is concluded that MENK could exert antitumor effect through precisely regulating opioid receptor mediated functions of DCs. In addition, MENK treated DCs may serve as a new immunotherapy approach against tumor.

The purpose of this study is to determine the efficacy of adjuvant immunotherapy with autologous cytokine-induced killer (CIK) for postoperative patients with gastric cancer and to investigate the impacts of the predictors on the efficacy of CIK immunotherapy. Ninety-two gastric cancer patients who have accepted radical resection were enrolled. The CIK and control groups were established by 1:1 matching on their baseline. As prognosis indicators, preoperative blood cell counts, the neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) were analyzed, respectively. Statistical analyses were done using IBM SPSS Statistics ver.19.0. CIK treatment significantly prolonged disease-free survival (DFS) (p

Cancer immunotherapy, due to its high anti-tumor efficacy, has attracted considerable attention globally from experts in various fields. However, immunotherapy could be severely toxic; not all patients may respond, thus requiring combination therapy. Therefore, a reasonable selection strategy for early treatment is urgently needed. It is vital to capture the dynamic, heterogeneous, and complex tumor behavior considering the absence of ideal companion biomarkers. Since tumor immune response involves tumor cells, several other cell types, and molecules in the tumor microenvironment, detection is very complex and variable. However, molecular imaging technology, namely the non-invasive whole-body molecular imaging by positron emission tomography and single-photon emission computed tomography, has shown considerable promise in tumor detection and cancer immunotherapy response. Identification of potential novel imaging biomarkers will allow a personalized treatment plan for every patient. Future imaging strategies for these molecules used alone or in combination or continuously, might help stratify patients before or during the early stages of immunotherapy, and might address the immunotherapy challenges encountered by the oncologists.

This article examined the associations between weight status (body mass index [BMI] and perceived weight measures) and school bullying experiences (being bullied and bullying others) of children and adolescents in urban China. A national representative sample ( n = 3,675) of all types of pre-college schools from seven Chinese cities was employed for the analysis. Students from all grades in high school, vocational school, middle school, and primary grades 4 to 6 were surveyed. Height, weight, perceived weight status, and experiences of being bullied and bullying others were obtained. The results demonstrated that the objective weight status (BMI) was neither associated with being bullied overall by others nor associated with bullying others, but the subjective/perceived weight status was significantly related to being bullied. Regarding the specific forms of being bullied, underweight (measured by BMI) students were less likely to be cyberbullied, whereas obese (measured by BMI) students were more likely to be cyberbullied. Students who perceived themselves as slightly fat and too fat were more likely to be verbally bullied. Overweight and obese students were more likely to bully others physically and relationally, and the students who perceived themselves as too thin were positively associated with physically bullying others. Moreover, BMI had a role in the involvement in school bullying for boys, whereas perceived weight status had a role in school bullying involvement in girls. These results indicate that school bullying prevention and intervention should consider weight status, as well as gender differences.

Tumor Necrosis Factor α (TNF-α), a multifunctional pro-inflammatory cytokine, is produced by macrophages/monocytes during acute inflammation, and plays a critical role in orchestrating the cytokine cascade in various inflammatory diseases. Previous studies demonstrated that TNF-α induces inflammatory responses in bone marrow mesenchymal stem cells (BMSCs) transplantation, leading to unsatisfactory effects and limit the clinical use of BMSCs. MicroRNAs are reported to involve in inflammation by regulating the expression of their targets in inflammatory response pathway. However, whether microRNAs mediate TNF-α-induced inflammatory responses in BMSCs remains elusive. Here, we found that TNF-α treatment induced an inflammatory response by increasing the levels of key inflammatory mediators, including IL-6, IL-1β, matrix metalloproteinase 9 (MMP9) and monocyte chemotactic protein-1 (MCP-1) in BMSCs. Moreover, real-time PCR result showed dramatically up-regulation of miR-128-3p after exposure to TNF-α. Interestingly, miR-128-3p over-expression exacerbated the TNF-α-induced inflammatory response, while suppression of miR-128-3p effectively eliminated the inflammatory response in BMSCs. Bioinformatic analysis identified sirtuin 1 is a direct target of miR-128-3p. Up-regulation of sirtuin 1 induced by resveratrol also diminished the TNF-α-induced inflammatory response in BMSCs. Altogether, our results indicated that miR-128-3p targets sirtuin 1 to mediate the TNF-α-induced inflammatory response in BMSCs, which may provide new strategies to protect against inflammatory-dependent impairments in BMSCs.

Background Non‐obstructive azoospermia (NOA), a serious phenotype of male spermatogenesis failure, is a multifactorial disease which is regulated by genetic, epigenetic, and environmental factors. Some gene structural variants have been demonstrated to be related to NOA. Loss‐of‐function mutations of KISS1R cause normosmic idiopathic hypogonadotropic hypogonadism (IHH) which result in azoospermia at the pre‐testicular level. The objective of this research was to investigate genetic variants of KISS1R in NOA patients. Methods The entire coding region of 52 spermatogenesis‐associated genes (KISS1R included) was sequenced from 200 NOA patients. Mutation screening was performed to identify genetic variations of these genes by targeted exome sequencing. Sequencing data analysis was carried out by a series of bioinformatics tools. Candidate variants confirmation was performed by Sanger sequencing. Functional analysis of candidate variants was evaluated using SIFT and PolyPhen‐2. Results Three heterozygous missense variants in KISS1R were identified in three patients, respectively. No deleterious variations in other candidate genes were found in the three patients. Two of these three variants, p.A211T and p.G186E, had been reported in the ExAC and dbSNP database, respectively, while the other variant p.A301D was novel. These variants were all predicted to be likely pathogenic by in silico analysis. Conclusion Our study revealed three heterozygous missense variants in KISS1R which expanded the mutation spectrum of KISS1R in infertile men with NOA in the northeast of China.

Rationale: This study aimed to report 1 family case with novel Y chromosome structural variations by an established next-generation sequencing (NGS) method using unique STSs. Patient concerns: The case studied was from a family with a father and son (the proband). G-band staining was used for karyotype analysis. Y chromosome microdeletions were detected by sequence-tagged site (STS)-PCR analysis and a new NGS screening strategy. Diagnoses: Semen analysis showed that the proband was azoospermic. The patient had an abnormal karyotype (45,X[48%]/46,XY[52%]). His father exhibited a normal karyotype. STS-PCR analysis showed that the proband had a deletion of the AZFb+c region, and his father had no deletion of STS markers examined. The sequencing method revealed that the patient had DNA sequence deletions from nt 20099846 to nt 28365090 (8.3 Mb), including the region from yel4 to the Yq terminal, and his father exhibited a deletion of b1/b3 and duplication of gr/gr. Interventions: The proband was advised to undergo genetic counseling, and consider the use of sperm from a sperm bank or adoption to become a father. Outcomes: The proband was azoospermic. AZFc partial deletions may produce a potential risk for large AZFb+c deletions or abnormal karyotypes causing spermatogenic failure in men. Lessons: The NGS method can be considered a clinical diagnostic tool to detect Y chromosome microdeletions. The partial AZFc deletions and/or duplications can be a risk of extensive deletions in offspring.

The purpose of this study was to investigate the prevalence of androgen receptor gene mutations in idiopathic male infertility in north-east China and to analyse the relationship between genotype and phenotype. Cohorts of 400 male patients with idiopathic infertility and 200 fertile controls were recruited. Clinical investigations were carried out by experienced andrologists. Targeted exome sequencing was used to detect androgen receptor gene mutations, and reproductive hormone levels were measured. We found four mutations in 8/400 patients (2%) and no mutations in controls. In addition, two recurrent mutations (p.S176R and p.A403V) were found in four patients and two patients respectively. Of four mutations, two (p.S233F and p.H715N) represented novel mutations. Almost all of the men with androgen receptor gene mutations showed smaller testicular volumes. However, the reproductive hormone levels were normal or lightly higher in seven men with mutation, apart from one man (P24) who had high follicle-stimulating hormone and luteinising hormone levels but a low testosterone level. In north-east China, there was a higher androgen receptor gene mutation rate among these infertile men, and the mutation of p.S176R deserved more attention particularly. Hormone levels and clinical phenotypes did not help in screening patients at risk of mutations.

Cell-based immunotherapy using natural killer (NK) cells, cytokine-induced killer (CIK) cells and dendritic cells (DCs) is emerging as a potential novel approach in the auxiliary treatment of a tumor. However, non-standard operation procedure, small-scale cell number, or human error may limit the clinical development of cell-based immunotherapy. To simplify clinical scale NK cells, CIK cells and DCs expansions, we investigated the use of the WAVE bioreactor, a closed system bioreactor that utilizes active perfusion to generate high cell numbers in minimal volumes. We developed an optimized rapid expansion protocol for the WAVE bioreactor that produces clinically relevant number of cells for our adoptive cell transfer clinical protocols. The high proliferative rate, surface phenotypes, and cytotoxicity of these immune cells, as well as the safety of cultivation were analyzed to illuminate the effect of WAVE bioreactor. The results demonstrated that the benefit of utilizing modern WAVE bioreactors in cancer immunotherapy was simple, safe, and flexible production.

School violence and bullying in China is under investigated, though it has become a national concern recently. Using updated national representative survey data collected in 2016 from seven provinces across China, covering students from all pre-college school types (primary, middle, high and vocational schools), this paper analyzes the prevalence of school bullying and the correlation with several school attributes. The incidences of reported bullying, bullying others and witnessing bullying are 26.10%, 9.03% and 28.90%, respectively. Primary school students are more likely to be involved in bullying behaviors. Students from elite schools (leading schools) are also more likely to be involved. Relation with teachers, relation with peers and perceived academic achievement are protective factors. Being a boy is the only significant predictor of school bullying among the family and demographic characteristics used. The results highlight the importance of school climate on preventing school violence and bullying, and a whole-school intervention approach is needed for future intervention.

Vacquinol‑1 (Vacq), a quinolone derivative, has recently been reported to display potent antitumor effects in glioblastomas by inducing cellular massive vacuolization and cell death. However, whether Vacq induces cytotoxicities in other types of cancers, and the potential underlying mechanism, remain to be investigated. In the present study, it was revealed that Vacq suppressed cell growth and colony formation in the hepatocellular carcinoma (HCC) cell lines BEL7402 and Huh7. In addition, treatment with Vacq increased the number of early and late apoptotic cells as assessed by flow cytometry with fluorescein isothiocyanate‑conjugated Annexin V and propidium iodide double staining. Notably, the effect by Vacq in the tested cells could be inhibited by pretreatment with a broad specificity caspase inhibitor Z‑VAD‑FMK, suggesting that Vacq may induce apoptosis in HCC cells. Morphologically, exposure to Vacq resulted in nuclear fragmentation and the apoptotic body formation in HCC cells. Furthermore, Vacq treatment increased the cleavage of caspase‑3, caspase‑9 and poly(adenosine diphosphate‑ribose) polymerase‑1. Mechanistic analysis revealed that Vacq upregulated the expressions of pro‑apoptotic proteins [B‑cell lymphoma 2 (bcl‑2)‑associated X protein (Bax) and Bcl‑2‑like protein 11] and downregulated the pro‑survival protein, Bcl‑2, expression in HCC cells. Furthermore, Vacq induced Bax translocation. Of note, Vacq displayed inhibitory effects on patient‑derived HCC cells in two‑dimensional (2D) and three-dimensional (3D) cultures. Taken together, the data suggested that Vacq induced intrinsic apoptosis and may be utilized as an effective reagent for HCC treatment.

The N-terminal truncated carboxypeptidase E (CPEΔN) protein is a novel biomarker of tumor metastasis. This study screened the H1299 cell line with a highly expressed CPEΔN gene for in vivo imaging experiment.Human CPEΔN gene was cloned into the luciferase lentiviral vector. H1299 cells transduced with CPEΔN or control lentiviral vectors were selected with 2 µg/mL puromycin. The expression of CPEΔN was identified through Western blot analysis, and luciferase activity was measured using luciferase reporters.The human CPEΔN lentiviral expression vector was successfully constructed. The transfection rate of H1299 cells by the lentivirus achieved 80%, with an infection multiplicity of 20. The H1299 cell line with high CPEΔN (H1299-CPEΔN) expression was established, with an increase in CPEΔN expression by four times compared with the control lentivirus-transfected H1299 cell line (H1299-control). As H1299-CPEΔN and H1299-control can effectively decompose luciferase substrates, they can be applied in in vivo imaging.H1299-CPEΔN and H1299-control can be used in in vivo imaging experiment for further research on molecular mechanisms and signal transduction to elucidate the role of CPEΔN in lung cancer metastasis.背景与目的 N端截短的羧肽酶E（N-terminal truncated carboxypeptidase E, CPEΔN）是一个新的肿瘤转移相关蛋白。本研究旨在筛选高表达CPEΔN的H1299肺癌细胞株，为完成小鼠活体成像实验创造条件。方法 构建CPEΔN的慢病毒表达载体。分别用CPEΔN慢病毒表达载体或对照慢病毒空载体转染H1299细胞，2 µg/mL的嘌呤霉素加压筛选。Western blot分析CPEΔN蛋白的表达，荧光素酶报告基因实验分析荧光素酶对底物的分解作用。 结果 当感染倍数（multiple of infection, MOI）是20时，慢病毒对H1299细胞的转染效率可以达到80%。CPEΔN高表达H1299细胞株（H1299-CPEΔN）和对照慢病毒载体表达H1299细胞株（H1299-control）中CPEΔN蛋白的表达量为4:1。H1299-CPEΔN和H1299-control均能够有效分解荧光素酶底物，可以满足活体成像实验的需求。结论 筛选出高表达CPEΔN的H1299肺癌细胞株，为活体成像实验的开展创造了条件，也为进一步解释CPEΔN促进肿瘤转移的分子机制奠定了基础。.

Background A retrospective study was conducted to evaluate the advantages of anterolateral thigh (ALT) flap in tongue reconstruction. Method From September 2008 to February 2012, patients receiving ALT flap tongue reconstruction were included in the study. Patients undergoing ALT flap were compared with those undergoing similar surgery with radial forearm flap (RFF). The medical records of the included patients were reviewed, and a questionnaire was used to assess acceptability of the surgery. Results All flaps (both ALT and RFF) were successful In the ALT group, most patients were satisfied with the appearance of the reconstructed tongue and the intelligibility of their speech, and there were fewer complications with this technique compared with the RFF. Conclusion The ALT flap is an ideal method for tongue reconstruction. The thickness and volume of the ALT flap can be adjusted based on the individual extent of the defect, and it can not only provide bulk but also ensure mobility, and it has other advantages also, including a long pedicle and low donor site morbidity.

A novel 3-5-dichlorosalicylaldehyde Schiff base chemodosimeter (compound 1) for water is designed and synthesized, and it works based on a water-triggered reaction of a Schiff base. Addition of trace amounts of water into 1 in various organic solvents leads to a fluorescence turn-on response and a simultaneous dual-channel signal modulation (both in the fluorescence and absorption spectra). Especially, 1 is found to be an outstanding fluorescence enhancement water sensor in methanol with an extremely low detection limit of 22 ppm. Consequently this probe can be utilized to detect trace water in commercial methanol. The quantitative detection of a wide range of water content is enhanced in THF and acetonitrile (0–35% v/v for THF and 0–20% v/v for acetonitrile), where the fluorescence peak intensity is nearly proportional to the amount of water added. Moreover, 1 can be used for monitoring pH through a novel ON–OFF–ON type signal modulation both in fluorescence and absorption spectra within a wide pH detection range. Thus, the chemodosimeter can not only be utilized to monitor the intracellular pH fluctuations, but also to accomplish simultaneous in situ staining of the cytosol and acidic organelles in two different channels, respectively.

Background Evidence demonstrated that sweat was an important factor affecting skin physiological properties. Aims We intended to assess the effects of exercise-induced sweating on the sebum, stratum corneum (SC) hydration and skin surface pH of facial skin. Materials & Methods 102 subjects (aged 5–60, divided into five groups) were enrolled to be measured by a combination device called ‘Derma Unit SSC3’ in their frontal and zygomatic regions when they were in a resting state (RS), at the beginning of sweating (BS), during excessive sweating (ES) and an hour after sweating (AS), respectively. Results Compared to the RS, SC hydration in both regions increased at the BS or during ES, and sebum increased at the BS but lower during ES. Compared to during ES, Sebum increased in AS but lower than RS. Compared to the RS, pH decreased in both regions at the BS in the majority of groups, and increased in frontal region during ES and in zygomatic region in the AS. There was an increase in pH in both regions during ES in the majority of groups compared to the BS, but a decrease in the AS compared to during ES. Discussion The study implies that even in summer, after we sweat excessively, lipid products should be applied locally in order to maintain stability of the barrier function of the SC. The study suggests that after a short term(1 h or less) of self adjustment, excessive sweat from moderate exercise will not impair the primary acidic surface pH of the facial skin. Conclusion Exercise-induced sweating significantly affected the skin physiological properties of facial region.