Objective: To construct a Family with sequence similarity 50 member A(FAM50A) gene knockout mouse insulinoma pancreatic β-cell line Beta-TC-6 using CRISPR/Cas9 gene editing technology and to prepare polyclonal antibodies specifically recognizing FAM50A. Methods: Two guide RNAs(sgRNAs) targeting the FAM50A gene were designed,and a recombinant plasmid expressing blue fluorescent protein(BFP) was constructed for gene knockout.The successfully constructed plasmid was transfected into Beta-TC-6 cells,and BFP-positive single cells were isolated for clonal expansion.The expanded monoclonal cell lines were genotyped by Sanger sequencing,and FAM50A protein expression was assessed by Western blot.Purified human recombinant FAM50A protein was used to immunize New Zealand rabbits for the preparation of a polyclonal antibody.The specificity of the prepared antibody was then validated using the successfully established FAM50A knockout cell line. Results: A monoclonal cell line with a successful knockout of the FAM50A gene was identified.Sanger sequencing confirmed base deletions at the target site.Western blot analysis showed a complete absence of FAM50A protein expression in this cell line.The prepared polyclonal antibody successfully recognized endogenous murine FAM50A protein in wild-type Beta-TC-6 cells and in hTERT-RPE1 cells overexpressing human FAM50A-GFP fusion protein,while no signal was detected in the FAM50A knockout cells. Conclusion This study successfully established a FAM50A gene knockout Beta-TC-6 cell model and generated a FAM50A polyclonal antibody,providing powerful tools for future research.

Objective:To explore the latent-classes of HIV high risk behaviors among college students, and the association between experiences of intimate partner violence (IPV) and HIV high risk behaviors, to provide evidence for reducing the HIV high risk behaviors among them.Methods:A cross-sectional study was conducted from October to December 2019 among university students from six higher education institutions in Zhuhai City, using a multi-stage cluster sampling method, with an estimated sample size of 1 318. The study included participants who self-reported being in a romantic relationship and having sexual experience within the past year. Data on sociodemographic characteristics, IPV experiences, and HIV high risk behaviors were collected. Latent-class analysis was performed on HIV high risk behaviors, and chi-squared tests and multivariable logistic regression were used to analyze the associations between IPV experiences and different latent classes of HIV high risk behaviors.Results:The effective response rate for the survey was 95.4% (12 235/12 821). 1 382 college students from Zhuhai were included as participants in the study, with 19.4% (268/1 382) self-reporting having experienced IPV. Latent-class analysis of HIV high risk behaviors classified the participants into three latent groups: low-risk group (78.1%, 1 079/1 382), multiple sexual partners/alcohol use before sex group (15.8%, 219/1 382), and high-risk group (6.1%, 84/1 382). Multivariable logistic regression analysis showed that students who had experienced psychological violence were more likely to be in the group that had multiple sexual partners/alcohol use before sex (a OR=2.51, 95% CI:1.48-4.27). Those who had experienced IPV (a OR=5.74, 95% CI:3.45-9.55), physical violence (a OR=9.26, 95% CI: 5.24-16.35), sexual violence (a OR=8.46, 95% CI:4.93-14.52), or psychological violence (a OR=15.99, 95% CI:8.64-29.57) were more likely to be in the high-risk group. Students who experienced two (a OR=9.37, 95% CI:3.55-24.71) or three types of IPV (a OR=50.09, 95% CI: 21.06-119.14) were more likely to be in the high-risk group compared to those with no IPV experiences. Conclusions:HIV high risk behaviors among college students in Zhuhai exhibited heterogeneity across different latent groups, and these groups have different associations with IPV experiences. Universities should tailor targeted HIV/AIDS education and prevention strategies based on the characteristics of each latent group to reduce HIV high risk behaviors among college students.

Objective:To explore the latent-classes of HIV high risk behaviors among college students, and the association between experiences of intimate partner violence (IPV) and HIV high risk behaviors, to provide evidence for reducing the HIV high risk behaviors among them.Methods:A cross-sectional study was conducted from October to December 2019 among university students from six higher education institutions in Zhuhai City, using a multi-stage cluster sampling method, with an estimated sample size of 1 318. The study included participants who self-reported being in a romantic relationship and having sexual experience within the past year. Data on sociodemographic characteristics, IPV experiences, and HIV high risk behaviors were collected. Latent-class analysis was performed on HIV high risk behaviors, and chi-squared tests and multivariable logistic regression were used to analyze the associations between IPV experiences and different latent classes of HIV high risk behaviors.Results:The effective response rate for the survey was 95.4% (12 235/12 821). 1 382 college students from Zhuhai were included as participants in the study, with 19.4% (268/1 382) self-reporting having experienced IPV. Latent-class analysis of HIV high risk behaviors classified the participants into three latent groups: low-risk group (78.1%, 1 079/1 382), multiple sexual partners/alcohol use before sex group (15.8%, 219/1 382), and high-risk group (6.1%, 84/1 382). Multivariable logistic regression analysis showed that students who had experienced psychological violence were more likely to be in the group that had multiple sexual partners/alcohol use before sex (a OR=2.51, 95% CI:1.48-4.27). Those who had experienced IPV (a OR=5.74, 95% CI:3.45-9.55), physical violence (a OR=9.26, 95% CI: 5.24-16.35), sexual violence (a OR=8.46, 95% CI:4.93-14.52), or psychological violence (a OR=15.99, 95% CI:8.64-29.57) were more likely to be in the high-risk group. Students who experienced two (a OR=9.37, 95% CI:3.55-24.71) or three types of IPV (a OR=50.09, 95% CI: 21.06-119.14) were more likely to be in the high-risk group compared to those with no IPV experiences. Conclusions:HIV high risk behaviors among college students in Zhuhai exhibited heterogeneity across different latent groups, and these groups have different associations with IPV experiences. Universities should tailor targeted HIV/AIDS education and prevention strategies based on the characteristics of each latent group to reduce HIV high risk behaviors among college students.

Objective·To analyze the anatomical and macroscopic characteristics of the pterygopalatine suture(PPS)in Chinese population using cone-beam computed tomography(CBCT)technology,and to preliminarily investigate its developmental pattern and its association with the correction of maxillary underdevelopment.Methods·A total of 134 CBCT images,taken from July to August 2023 at Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,were categorized into six age groups.The PPS landmarks were utilized to determine the overall transverse position(XPPS),sagittal position(YPPS),insertion angle(IAP),insertion width of the pyramidal process(IWP),and insertion depth of the pyramidal process(IDP).The pterygomaxillary junction was identified through multiplanar observations.Regression analysis was performed to assess the correlation of these parameters with age and gender,and pairwise comparisons were made to determine the stable age range for parameter changes.Paired t-tests and paired chi-squared tests were conducted to analyze the bilateral parameter differences.Results·Results showed that YPPS and IDP did not exhibit significant correlation with age,whereas XPPS,IAP,IWP,and the incidence of pterygomaxillary fusion were significantly positively correlated with age(P<0.01).Gender differences were only significant for XPPS,with males demonstrating greater values than females(P<0.01).Analysis of age-related trends indicated significant differences in XPPS between group 1(6 years≤age<9 years)and group 2(9 years≤age<12 years)(female:P=0.006,male:P=0.004);significant differences in IAP were observed between group 2 and group 3(12 years≤age<15 years)(P=0.042),with 98.5%of samples having an IAP greater than 45 degrees;IWP differences were significant between group 1 and group 3(P=0.016),and the pterygomaxillary fusion incidence was significantly different among group 1,2,and 3(group 1 vs.2:P<0.001,group 2 vs 3:P=0.037,group 1 vs 3:P<0.001),with an incidence rate exceeding 90%in adults.No significant bilateral differences were found for all parameters.Conclusion·No significant changes were observed in YPPS and IDP after the age of 6,indicating a trend towards fusion of the maxilla with the pterygoid process.The overall transverse position of the PPS tends to stabilize around 12 years of age,while the IAP and IWP continue to increase and reach stability around 15 years of age,with a sagittal insertion orientation of the pyramidal process into the pterygoid notch.

Diabetes, hypertension, and dyslipidemia, collectively referred to the " Three Highs, " represent increasingly prevalent metabolic risk factors in China. Many individuals experience all three conditions concurrently, significantly heightening the risk of cardiovascular disease and mortality. Although the National Metabolic Management Center(MMC) has been established for over eight years and has its unique features, the awareness, treatment, and control rates of these diseases in China remain low, and the efficiency of community management is insufficient. According to the previous two editions of management guidelines and the most recent domestic and international diagnostic and treatment guidelines, this paper conducts an in-depth analysis of the operational experience and management strategies of the MMC. Its aim is to improve the efficiency of grassroots MMC mode management for " Three Highs" patients and ensure that patients receive more standardized management.

Diabetes, hypertension, and dyslipidemia, collectively referred to the " Three Highs, " represent increasingly prevalent metabolic risk factors in China. Many individuals experience all three conditions concurrently, significantly heightening the risk of cardiovascular disease and mortality. Although the National Metabolic Management Center(MMC) has been established for over eight years and has its unique features, the awareness, treatment, and control rates of these diseases in China remain low, and the efficiency of community management is insufficient. According to the previous two editions of management guidelines and the most recent domestic and international diagnostic and treatment guidelines, this paper conducts an in-depth analysis of the operational experience and management strategies of the MMC. Its aim is to improve the efficiency of grassroots MMC mode management for " Three Highs" patients and ensure that patients receive more standardized management.

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.

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 .