Objective To construct a lentiviral vector over-expressing β-catenin gene by multisite Gateway technology and confirm its effect.Methods By using multisite Gateway clone technique,the entry clone of pDown-Ctnnb1 was constructed using BP recombination reaction.Then,LR recombination reaction was performed among pUp-EF1A,pDown-Ctnnb1,pTail-IRES/DsRed-Express2 and pLV.Des3d.P/puro to generate an expression clone of pLV.EX3d.P/puro-EF1A＞Ctnnb1 ＞IRES/DsRed-Express2.In each step,PCR and sequencing analysis were used to verify the constructions.When it was verified that plasmids were transfected into 293T cells,PT-PCR was performed to determine the mRNA level of β-catenin gene.Results Both PCR and sequencing analysis revealed that β-catenin over-expression gene was inserted into the target site and the insertion sequence was perfectly corrected.The RT-PCR results showed that the expression of β-catenin gene was significantly upregulated.Conclusion The lenvivirus-mediate β-catenin over-expression gene was successfully constructed..

Objective To design pedometer shoes through wireless transmission step calculation.Methods The transmitting terminals in the soles acquired the pressure signals,and then the signals were transmitted to the SCM after conversion,amplification and comparison.AT89S52 SCM was used for step calculation and then the results were sent to the receiver through nRF905.Results The receiver had three modes,such as the step-calculation mode with the time and step calculation result displayed,the running mode with the distance displayed and the function of stopwatch,and the prompt mode with the distance and consumed power displayed.Conclusion The wireless embedded pedometer shoes are multifunctional,easy to operate and reliable.

AIM: To compare the cytotoxic effect of evodiamine with chemotherapy drugs on A375-S2 cells, and to examine the relationship between the effects of PKC and ERK on evodiamine-induced cell death. METHODS: MTT assay and Western blot analysis were applied. RESULTS: Compared to actinomycin D, cisplatin and 5-FU, evodiamine showed less cytotoxic effects on A375-S2 cells, but it induced more significant inhibition of proliferation in A375-S2 cells incubated with evodiamine for 24 h, followed by continuous culture in drug-free medium. The activation of PKC induced by 10 ?g?L -1 PMA partially blocked evodiamine-induced cell death, which was reversed by PKC and ERK inhibitors. Moreover, evodiamine down-regulated the expressions of ERK and phosphorylated ERK. CONCLUSION: Evodiamine has a strong inhibitory influence on proliferation of A375-S2 cells, even after removal of evodiamine. Evodiamine blocks the protective role of ERK to A375-S2 cells through the downregulation of ERK and phosphorylated ERK expression. [

Objective:To investigate the current situation and correlation of perceived social support,achievement motivation,and depression among nursing graduates of a medical college in Anhui Province.Methods:A convenient sampling method was used to select 286 graduates from the School of Nursing in a medical college in Anhui Province as the research subjects.They were investigated using a general information questionnaire,the Perceived Social Support Scale,the Achievement Motivation Scale,and the Self-Rating Depression Scale.SPSS 23.0 and SPSS process plug-in were used to conduct correlation analysis and mediation effect test.Results:Perceived social support and achievement motivation were negatively correlated with depression(r=-0.413,-0.306,P＜0.01).Perceived social support was positively correlated with achievement motivation(r=0.323,P＜0.01).Achievement motivation partially mediated the relationship between perceived social support and depression,with a mediating effect of 14.99%of the total effect.Conclusion:The perceived social support among nursing graduates can have a direct impact on depression,and can also predict depression through the indirect effect of achievement motivation.

Objective To establish ovarian cancer cell line SKVO3 that can stably express human ADP ribosylation factor-4 (ARF4). Methods A eukaryotic expression vector pCDH-CMV-MCS-EF1-Puro/ARF4 was constructed and transfected into SKOV3 cells after verifying by DNA sequencing. The expression of ARF4 mRNA was verified by real-time quantitative PCR (qRT-PCR). Then, the recombinant plasmid with lentiviral packaging plasmids were co-transfected into SKOV3 cells for packaging. The recombinant lentiviral particles LV-ARF4 were collected and transfected into SKOV3 cells, and the stable transfected SKOV3 cell line was screening by culture with puromycin. The expression of ARF4 gene was detected by qRT-PCR and Western Blot. Results A eukaryotic expression vector pCDH-CMV-MCS-EF1-Puro/ARF4 was successfully constructed. The vector could significantly up-regulate the expression of ARF4 mRNA in SKOV3 cells and be successfully packaged into recombinant lentiviral particles in HEK-293T cells. Compared with the control group, the relative expression level of ARF4 mRNA and protein in SKOV3 cells was significantly increased after the infection with LV-ARF4 (all P<0.001). Conclusion The recombinant plasmid pCDH-CMV-MCS-EF1-Puro/ARF4 and lentiviral vector LV-ARF4 were successfully constructed. The establishment of stably infected SKOV3 cell line with LV-ARF4 provides an experimental foundation for further studies on the biological function of ARF4 in ovarian cancer.

Objective To study the effect of citicoline on mild cognitive impairment MCI) in pa tients with middle cerebral artery stenosis (MCAS).Methods Eighty-six MCAS patients with MCI were divided into citicoline group (n=44) and control group (n=42).The patients in citicoline group were treated with citicoline (0.2 g,3 times a day) for 6 months on the basis of conventional treatment.Cerebrovascular reserve,PI,BHI and Vm between the two groups were compared by transcranial Doppler ultrasonography after treatment.MCI between the two groups was assessed according to the MoCA after treatment.Results The rate of cerebrovascular reserve,PI,BHI and Vm were significantly higher in citicoline group than in control group (13.59％± 1.16％ vs 7.61％±1.12％,P＜0.01;0.51±0.16 vs 0.58±0.12,P＜0.05;1.36±0.08 vs 0.74±0.11,P＜ 0.01;32.63％ ±2.32％ vs 16.92％ ± 1.68％,P＜0.05).The total MoCA score,attention,language,visuospatial and executive function,abstract,naming,orientation and memory were significantly higher in citicoline group than in control group (P＜0.01).Conclusion Early citicoline treatment can improve cerebrovascular reserve and alleviate MCI in MCAS patients.

To explore the clinical value of contrast-enhanced ultrasound (CEUS) in differentiating benign and malignant focal liver lesions (FLLs) with SonoVue, CEUS was used to examine 113 pa- tients with focal liver lesions (FLLs) in our hospital during July 2005 to December 2006. All the pa- tients underwent contrast-enhanced CT (CECT) or contrast-enhanced MRI(CEMRI). Except for pa- tients with focal fatty sparings (n=18) and with hemangiomas (n=8), all the patients were confirmed by operation or ultrasonic-guided liver puncture biopsy. A sulfur hexafluoride gas-based contrast agent was used with a MI of 0.15 to 0.17. Forty-eight cases of malignant FLLs, including 30 hepato- cellular carcinomas (HCCs), 2 cholangiocarcinomas and 16 metastatic tumors, were detected. Sev- enty-eight cases of benign FLLs, including 33 hemangiomas, 9 focal nodular hyperplasias (FNHs), 19 focal fatty sparings, 5 abscesses, 7 regenerative nodules and 2 inflammatory pseudo-tumor, were in- volved. The contrast pattern of benign and malignant FLLs was quite different. CEUS has higher specificity and sensitivity than conventional ultrasound in differentiating benign and malignant FLLs.

Objective:To analyze clinical features, short-term efficacy and side effects of salvage re-irradiation therapy for patients with locally recurrent esophageal cancer after definitive chemoradiotherapy, to investigate the prognostic factors of re-irradiation with precise radiotherapy techniques.Methods:A retrospective analysis was performed on patients with locally recurrent esophageal squamous cell carcinoma after definitive chemoradiotherapy treated in the Fourth Hospital of Hebei Medical University from January 2008 to December 2016. The patients underwent re-irradiation therapy (re-RT) or re-irradiation therapy concurrent chemotherapy (re-CCRT). The main observation index was after-recurrence survival (ARS), which was calculated by Kaplan-Meier method for survival analysis. Univariate analysis was conducted by log-rank test, and multivariate analysis was performed by Cox regression model.Results:A total of 109 patients were included, with a median age of 66 years (43-89 years), and a median follow-up time of 120.8 months (79.0-176.5 months). The objective response rates (ORR) and dysphagia improvement rates (DIR) in all patients were 64.2% and 63.0%, respectively. The median ARS and 1-, 3-, 5-, 8-year survival rates in all patients were 7.8 months and 32.1%, 9.2%, 7.3% and 2.3%, respectively. The median ARS and 1-, 3-, 5-years survival rates were 10.8 months and 45.9%, 13.5%, 10.8% for patients with time to recurrence (TTR) ≥24 months, significantly longer than those of 5.7 months and 25.0%, 6.9%, 5.6% for patients with TTR<24 months ( χ2=7.99, P=0.005). The median ARS in groups with re-irradiation dose of ≤50 Gy,>50-54 Gy, and>54 Gy groups were 5.7, 10.0 and 8.1 months, respectively ( χ2=6.94, P=0.031). The 1-, 3- and 5-year survival rates were 30.4%, 5.1%, and 3.8% for re-RT versus 36.7%, 20.0%, and 16.7% for re-CCRT ( χ2=2.12, P=0.145). Multivariate analysis showed that TTR ( HR=0.607, 95% CI=0.372-0.991, P=0.046) and lesion length ( HR=0.603, 95% CI=0.371-0.982, P=0.042) were the independent factors for ARS. There was no significant difference in ≥2 grade pneumonitis and 2-3 grade radiation esophagitis between the re-RT and re-CCRT groups ( χ2=0.25, P=0.619; χ2=0.51, P=0.808). The morbidity of ≥2 grade myelosuppression in the re-RT group was significantly lower than that in the re-CCRT group (3.7% vs. 36.7%, χ2=18.15, P<0.001). Conclusions:Precise re-irradiation therapy for patients with locally recurrent esophageal cancer after definitive chemoradiotherapy can alleviate dysphagia, but ARS remains poor. Re-irradiation dose range from>50-54 Gy may be suitable for locally relapse patients as salvage treatment. Patients with TTR≥24 months and lesion length ≤5 cm obtain favorable prognosis.

Objective:To explore the effects of magnanimous therapy on the magnanimous and enterprising traits of lung cancer patients and the analysis of related factors.Methods:Totally 197 patients with lung cancer were divided into individual group ( n=62), team group ( n=75) and control group ( n=60). Comparison and correlation analysis were applied to the data before and after the electroencephalogram and the magnanimous questionnaire, the cancer response questionnaire, the T-type psychological scale, the cancer heart state questionnaire and the cancer patient's life function index scale. t test, analysis of variance and Pearson correlation analysis were processed by SPSS 23.0. Results:After treatment, the " enterprising" dimension and " magnanimous" dimension of individual group and the " enterprising" dimension of the team group ((3.035±0.309), (3.041±0.265), (3.173±0.371)) were higher than that before treatment((2.934±0.326), (2.908±0.315), (3.130±0.387), all P<0.05). There was negative correlation between " magnanimous" dimension of the magnanimous questionnaire and " subconscious" dimension of the T-type psychological scale in individual group( r=-0.280, P<0.05). In team group, the " enterprising" dimension of the magnanimous questionnaire was negatively correlated with " Psychological" and " Yield" dimension of the cancer heart state questionnaire( r=-0.279, -0.285, P<0.05), and positively correlated with " Facing" of the cancer response questionnaire, " Good physical condition and ability" and " Psychological well-being" dimension of the cancer patient's life function index scale( r=0.367, 0.402, 0.379, P<0.05). There was a negative correlation between the " enterprising" dimension of the magnanimous questionnaire and the beta wave value in individual group. Conclusion:The magnanimous therapy can improve enterprising and magnanimous level of patients with lung cancer, and the effects are related with the above-mentioned psychosomatic factors.

Objective:To explore changes in high-risk sexual behaviors and associated factors in HIV-infected men who have sex with men (MSM) in industrial workers, and provide evidence for designing behavioral interventions for this population.Methods:In this observational study, HIV-infected MSM were recruited in industrial workers using convenient sampling during August to September 2021. The sample size was estimated to be 530. A questionnaire was used and combined with routine follow-up to collect socio-demographic characteristics, high-risk sexual behaviors, partner notification, viral load testing and history of sexually transmitted diseases before and after diagnosis of HIV infection. The χ2 test was used to analyze the changes in high-risk sexual behaviors before and after diagnosis and logistic regression was conducted to identify factors associated with high-risk sexual behaviors. Results:A total of 560 HIV-infected MSM in industrial workers were recruited in this study. Of whom, 32.1% (180/560) had unprotected anal intercourse (UAI) within 12 months after diagnosis . The proportions of those having UAI with casual, commercial and regular same-sex partners significantly decreased from 73.4% (381/519), 75.1% (187/249) and 69.5% (207/298) within 12 months before diagnosis to 36.2% (146/403), 40.2% (86/214) and 34.2% (67/196) within 12 months after diagnosis , respectively. Educational level of college or above (a OR=0.41, 95% CI:0.23-0.71), passive anal sex (a OR=0.40, 95% CI:0.19-0.85), both active and passive anal sex after diagnosis (a OR=0.40, 95% CI:0.20-0.83) and no unprotected oral sex (a OR=0.02, 95% CI:0.01-0.05) were negatively associated with UAI within 12 months after diagnosis. Whereas, not considering necessary to use condom consistently after having repeated undetectable viral load (a OR=3.02, 95% CI:1.37-6.69) was positively associated with UAI within 12 months after diagnosis. Conclusions:Compared with that before diagnosis of HIV infection, although the prevalence of UAI seemed to decrease in HIV-infected MSM in industrial workers after diagnosis, nearly one third of them had high-risk sexual behaviors. Therefore, relevant interventions should be strengthened to reduce high-risk sexual behaviors.