[Objective] To investigate the variation of Q promoter (Qp) in nasopharyngeal carcinoma (NPC) cells, and to compare the existing two mutant sites [62 225 site(g→a)and 62 422 site (g→c) ] Qp in NPC cells with the Qp in B95.8 cell line in the functional and biological difference. [Methods] The Qp sequence was amplified in the samples from 29 cases of paraffin-embedded tissues of NPC suffers and 14 cases of peripheral blood of healthy adults by polymerase chain reaction (PCR) method (totally 43 cases). The point mutations on specified sites were analyzed and statistically compared from sequencing results. The sequences of variant and prototype Qp were amplified by PCR and cloned into luciferase reporter vector (pGL3-basic), then transfected into HaCat cells respectively. The transcriptional activity was compared between variant and prototype Qp using luciferase reporter system. The DNA binding affinity of mutant and prototype Qp to Sp1 was compared through chromatin immunoprecipitation (CHIP) method since mutation of nt 62 225 located in a Spl binding site. [Results] The mutation rate of Qp was significantly higher in NPC compared with healthy controls (P=0.039 5, <0.05), which suggested the variant Qp was closely associated with NPC. The transcription of the luciferase gene promoted by variant Qp was significant more than that of prototype Qp in transient transfection assay (2.5:1, P<0.05). The binding affinity of variant Qp to Sp1 was about 1.52 times higher than that of prototype Qp as determined by quantitative ChIP assay. [Conclusions] The transcriptional activity was enhanced in variant Qp in NPC cells compared with prototype, which possibly through the higher binding affinity to Sp1. We suggest that the mutated Qp may play an important role during the EBV infection and transformation of nasopharyngeal epithelium.

Objective:To investigate the functional status of dendritic cells(DCs)in oral leukoplakia(OLK)tissues.Methods:The expression of DC-specific markers CD1 a,CD209,CD1 23 and CD83 in 20 cases of OLK with abnormal dysplasia,1 0 with simple dys-plasia and 1 0 of normal oral mucosa tissues was examined by immunohistochemistry.Results:CD1 a and CD209 positive DCs were found in all cases.More CD1 a positive Langehans cells(LCs)in lamina propria were found in OLK with abnormal dysplasia than in normal o-ral mucosa and OLK with simple dysplasia(P <0.01 ).A great mount of CD209 positive stromal DCs were recruited in OLK.There was no CD83 positive and CD1 23 positive cell in normal oral mucosa,however,CD83 positive mature DCs and CD1 23 positive plasmacytoid DCs(PDCs)were observed in OLK(P <0.01 ).Conclusion:OLK is characterized by the recruitment of different subsets of DCs,the different DC subsets may play an important role in the development of OLK.

Objective To investigate the relationships between aspirin resistance (AR) and laboratory indexes and different types of traditional Chinese Medicine(TCM)syndrome in patients with acute cerebral infarction. Methods Two hundred and eight different types of TCM syndromes of patients with acute cerebral infarction admitted from January 2012 to November 2013 in the Neurology Department of Shanghai Seventh People's Hospital were divided into AR group and aspirin sensitive(AS)group according to the rate of AR. Simultaneously,28 healthy volunteers in the same period were assigned in a healthy control group. The changes of red blood cell volume distribution width coefficient of variation(RDW-CV),platelet count(PLT)and homocysteine(Hcy)levels were observed in the three groups. The correlation between different types of TCM syndromes and AR,PLT,RDW-CV,and Hcy was analyzed by non-conditional logistic regression. Results The total incidence of AR was 29.32%(61/208)in 208 patients with acute cerebral infarction. There were 165 cases with Qi deficiency and blood stasis syndrome,the incidence of AR being 26.06%(43/165);32 cases with wind phlegm obstructing channel syndrome,the incidence of AR, 43.75%(14/32);11 cases with liver yang hyperactivity syndrome,the incidence of AR,36.36%(4/11);in the comparisons,the incidence rates of AR among the above types of syndromes had no statistical significant differences (P>0.05). Compared with the healthy control group,the levels of PLT,RDW-CV,Hcy in AR group and AS group of various types of TCM syndrome were increased,the PLT and RDW-CV levels in patients with Qi deficiency and blood stasis syndrome in AR group were more significantly elevated in the comparisons between AR and AS groups, there were statistical significant differences〔PLT (×109/L):212.16±66.48 vs. 187.54±56.85, RDW-CV:(14.34±3.16)% vs.(13.20±2.16)%,both P<0.05〕;the level of Hcy in patients with wind phlegm obstructing channel syndrome in AR group was increased more significantly than that in AS group,the difference between the two groups being statistically significant(μmol/L:27.29±18.64 vs. 21.36±14.61,P<0.05). Logistic regression analysis showed,increased PLT〔odds ratio(OR)=1.007 2,95%confidence interval(CI):1.001 2-1.013 2,P=0.018 5〕and RDW-CV〔OR=1.165 4,95%CI:1.007 9-1.347 4,P=0.038 8)was independence risk factor of AS development. Conclusion The elevation of RDW-CV,PLT,Hcy in level reflects the index of AR production, especially in patients with acute cerebral infarction accompanied by Qi deficiency and blood stasis syndrome and wind phlegm obstructing channel syndrome.

Objective:To explore the efficient construction of HSF1 gene knockout mouse model using CRISPR/Cas9 gene editing technology, and to establish the early basis for the mouse model of primary osteosarcoma.Methods:According to exon 9 of HSF1 gene structure, the corresponding GRNA (guideRNA) was selected and screened. Then the transcription template of sgRNA (small guide RNA) was amplified by PCR, and four up stream primers were obtained. Subsequently, sgRNA was transcribed in vitro and screened by Tube Screen platform to screen the sgRNA with effective cutting, and the sgRNA with the highest cutting efficiency was selected from the screening results for subsequent experiments. The transcription template of SPCas9mRNA was amplified by PCR, and then Cas9mRNA was transcribed in vitro. The sgRNA transcribed in vitro and Cas9mRNA were injected into the fertilized eggs of healthy C57BL/6 mice, and the tissue was extracted from the tail of the born mice and identified by PCR sequencing. Heterozygous female mice of F0 generation were selected to mate with wild-type male mice too btain F1 generation off spring. The mutation of gene bases of F1 generation mice was detected by AGAR gel electrophoresis and gene sequencing. The heterozygous male mice of the F1 generation and female mice of the F0 generation were back crossed to obtain the F2 generation daughter mice. The tail tissues were cut and sequenced to obtain the F2 generation homozygous knockout mice. PCR was used to observe the cutting efficiency of sgRNA and the sequencing of rat tail tissue, and SNAPGene software was used for gene sequence alignment to determine the deletion of base fragments.Results:The up stream primers sgRNA-1 Primer-f, sgRNA-2 Primer-f, sgRNA-3 Primer-f, sgRNA-4 Primer-f and down stream primers sgRNA-4 Primer -r were obtained by PCR amplification. After in vitro tran scription and screening of sgrRNA, sgrRNA-1, sgrRNA-2 and sgrRNA-4 had high cleavage efficiency and were selected for subsequent experiments. T7 promoter was added to the 5 'end of Cas9 mRNA, and Cas9 mRNA was obtained by PCR and in vitro transcription kit. Mixed Cas9-sgRNA solution was injected into the fertilized eggs of mice and cultured. The cultured two-cell fertilized eggs were injected into the ampulla of the pseudo pregnant female mice, and the F0 generation mice were obtained successfully. A total of 8 heterozygous mice of F0 generation were obtained by Agar gel electrophoresis. Three heterozygous knockout mice of F1 generation were obtained by breeding the female heterozygous mice of F0 generation with healthy wild-type male mice and PCR and sequencing. Three heterozygous male mice of F1 generation were back crossed with female mice of F0 generation 3 to obtain F2 generation mice. Through the observation of electrophoresis and sequencing results of F2 generation mice, it was confirmed that 7 mice were missing HSF1 base sequence, and the electrophoresis results showed mutant bands and no wild-type bands, which were identified as homozygous. The F2 generation homozygous mice were able to breed stably. As eries of results proved that the HSF1 gene knockout mouse model was successfully established in this experiment.Conclusion:CRISPR/Cas9 technology was successfully used to construct HSF1 gene knockout mouse model, with strong stability and high reproducibility, which laida foundation for further study of HSF1 gene expression products and establishment of mouse model of primary osteosarcoma.

Objective To investigate the positive practice environments ( PPE ) of nurses and influencing factors at public hospitals , for reference of building a better PPE .Methods A national cross-sectional survey was performed at 77 public hospitals across seven provinces/metropolises, involving 5374 nurses.PPE included organizational management (internal) and nurses-patient relationship (external). Results The scoring of positive practice environment was 18.51 ±4.69 (total score of 40).The scoring of organizational management and nurses-patient relationship was 9.87 ±3.11 and 8.64 ±2.51 respectively. The scoring of PPE of nurses of general hospital ( GH) was higher than that of traditional Chinese medicine hospital(TCMH) (18.68 ±4.68 versus 18.08 ±4.67, P<0.01).Multivariable analysis showed that , compared with nurses who had not very much pressure about performance assessment , the scores of those who had were declined (βGH =-1.15, 95％CI: -1.55 to -0.76;βTCMH =-1.29, 95％CI: -1.92 to-0.66 ) );compared with nurses who paid less efforts in communicating with their patients , the scoring of those with greater efforts was higher (βGH =2.43, 95％CI:2.00 to 2.86;βTCMH =2.84, 95％CI:2.19 to 3.49).Conclusions PPE of nurses is poor in general in China , and the externally stressful nurse-patient relationship deserves greater attention and efforts than inefficient organization management internally .To improve PPE of nurses , hospitals need to moderate nurses′performance assessment and the nurses need to pay more attention to patient communication .

Objective The changes and risk factors of FeNO, CRP and PCT in patients with bronchial asthma complicated with sleep apnea syndrome (SAS) in Shanghai area were analyzed to provide theoretical basis for the prevention and treatment of SAS in patients with bronchial asthma. Methods A total of 436 patients with bronchial asthma admitted to our hospital from January 2019 to June 2022 were selected and divided into control group and experimental group according to whether SAS occurred during hospitalization. The experimental group was divided into three subgroups according to the apnea hypopnea index (AHI) values: mild group (AHI>15 times /h or <30 times /h), moderate group (AHI>15 times /h or <30 times /h) and severe group (AHI≥30 times /h).The serum FeNO, CRP and PCT levels were compared between the two groups. Clinical data of patients were collected from the medical record system. Univariate analysis and logistic regression were used to analyze the factors affecting the occurrence of SAS in patients with bronchial asthma. Results Among the 436 patients with bronchial asthma,158(36.24%)had SAS,including 76 mild,54 moderate and 28 severe;The levels of serum FeNO,CRP,PCT and AHI in the experimental group were significantly higher than those in the control group(P<0.05).Serum FeNO,CRP,PCT and AHI levels in severe group were significantly higher than those in mild group(t=5.830,7.597,7.885,40.684,P<0.05).Serum FeNO,CRP,PCT and AHI levels in severe group were significantly higher than those in moderate group(t=2.895,3.455,3.001,18.841,P<0.05).The levels of serum FeNO,CRP,PCT and AHI in the moderate group were significantly higher than those in the mild group(t=2.924,4.157,6.201,23.640,P<0.05).BMI>28 kg/m²(OR=5.629),allergic rhinitis(OR=6.166)and neck circumference>40 cm(OR=5.265)were independent risk factors for SAS in patients with bronchial asthma(P<0.05);Pearson correlation analysis showed that AHI was positively correlated with serum FeNO,CRP and PCT levels in patients with bronchial asthma and SAS(r=0.471,0.436,0.502,P<0.05). Conclusions Patients with old bronchial asthma in Shanghai area have a higher risk of SAS, and the severity of the disease is positively correlated with the levels of FeNO, CRP and PCT. In particular, obese patients with allergic rhinitis should be given early intervention to reduce the risk of SAS.

As known, the benefits of photothermal therapy (PTT) are greatly limited by the heat tolerance of cancer cells resulting from overexpressed heat shock proteins (HSPs). Then HSPs further trigger the formation of stress granules (SGs) that regulate protein expression and cell viability under various stress conditions. Inhibition of SG formation can sensitize tumor cells to PTT. Herein, we developed PEGylated pH (low) insertion peptide (PEG-pHLIP)-modified hollow copper sulfide nanoparticles (HCuS NPs) encapsulating the SG inhibitor ISRIB, with the phase-change material lauric acid (LA) as a gate-keeper, to construct a pH-driven and NIR photo-responsive controlled smart drug delivery system (IL@H-PP). The nanomedicine could specifically target slightly acidic tumor sites. Upon irradiation, IL@H-PP realized PTT, and the light-controlled release of ISRIB could effectively inhibit the formation of PTT-induced SG to sensitize tumor cells to PTT, thereby increasing the antitumor effect and inducing potent immunogenic cell death (ICD). Moreover, IL@H-PP could promote the production of reactive oxygen species (ROS) by tumor-associated macrophages (TAMs), repolarizing them towards the M1 phenotype and remodeling the immunosuppressive microenvironment. In vitro/vivo results revealed the potential of PTT combined with SG inhibitors, which provides a new paradigm for antitumor and anti-metastases.