Objective:ANXA2 plays a crucial role in cancer metastasis, but its mechanism is not yet fully understood. Therefore, it is necessary to establish an ANXA2 gene knockout mouse model to provide an effective tool for subsequent studies on ANXA2-related mechanisms.Methods:A gene knockout mouse model was constructed using CRISPR/Cas9 technology. The model was validated through tissue DNA extraction followed by polymerase chain reaction (PCR), sequencing, and western blot to confirm ANXA2 genotype and protein expression. The successfully constructed models were divided into a model group and a wild-type (WT) group for the creation of a mouse tail vein injection Lewis lung carcinoma (LLC) metastasis model. Metastatic foci formation was monitored using in vivo imaging technology, and the survival rates of the two groups were compared. Results:An sgRNA sequence targeting the first exon of ANXA2 was designed, and 16 founder mice were obtained through microinjection. Through consanguineous hybridization, 30 homozygous offspring were ultimately acquired. After establishing the strains of the mouse model, mice were divided into the ANXA2 knockout group and the WT group, with 8 mice in each group. An LLC lung metastasis model was established in both groups. Compared with the WT group, the number of metastatic foci was significantly increased in the ANXA2 knockout group (7 vs. 1), and the fluorescence intensity was stronger in the WT group than in the knockout group ( P=0.002). Using the GEPIA2 database to analyze ANXA2 gene expression in tumor tissues and normal tissues of lung cancer patients, it was found that ANXA2 expression levels were significantly higher in lung cancer tumor tissues compared to normal tissues ( P＜0.05). The database included data from 478 lung cancer patients, and patients were stratified into high-expression and low-expression groups based on ANXA2 levels. Compared to the low-expression group, patients in the high-expression group exhibited significantly shorter disease-free survival and overall survival ( P＜0.05, respectively). The survival time of mice in the ANXA2 knockout group (median survival time, 43 days) was significantly longer compared to the WT group (median survival time, 26 days; P=0.017). Additionally, ANXA2 expression is significantly associated with the prognosis of lung cancer patients ( P=6.4e-14). Conclusions:ANXA2 is closely associated with cancer metastasis and holds potential as a new target for metastasis treatment. Further in-depth research will greatly facilitate the transition of ANXA2 from basic research to clinical application.

Objective:ANXA2 plays a crucial role in cancer metastasis, but its mechanism is not yet fully understood. Therefore, it is necessary to establish an ANXA2 gene knockout mouse model to provide an effective tool for subsequent studies on ANXA2-related mechanisms.Methods:A gene knockout mouse model was constructed using CRISPR/Cas9 technology. The model was validated through tissue DNA extraction followed by polymerase chain reaction (PCR), sequencing, and western blot to confirm ANXA2 genotype and protein expression. The successfully constructed models were divided into a model group and a wild-type (WT) group for the creation of a mouse tail vein injection Lewis lung carcinoma (LLC) metastasis model. Metastatic foci formation was monitored using in vivo imaging technology, and the survival rates of the two groups were compared. Results:An sgRNA sequence targeting the first exon of ANXA2 was designed, and 16 founder mice were obtained through microinjection. Through consanguineous hybridization, 30 homozygous offspring were ultimately acquired. After establishing the strains of the mouse model, mice were divided into the ANXA2 knockout group and the WT group, with 8 mice in each group. An LLC lung metastasis model was established in both groups. Compared with the WT group, the number of metastatic foci was significantly increased in the ANXA2 knockout group (7 vs. 1), and the fluorescence intensity was stronger in the WT group than in the knockout group ( P=0.002). Using the GEPIA2 database to analyze ANXA2 gene expression in tumor tissues and normal tissues of lung cancer patients, it was found that ANXA2 expression levels were significantly higher in lung cancer tumor tissues compared to normal tissues ( P＜0.05). The database included data from 478 lung cancer patients, and patients were stratified into high-expression and low-expression groups based on ANXA2 levels. Compared to the low-expression group, patients in the high-expression group exhibited significantly shorter disease-free survival and overall survival ( P＜0.05, respectively). The survival time of mice in the ANXA2 knockout group (median survival time, 43 days) was significantly longer compared to the WT group (median survival time, 26 days; P=0.017). Additionally, ANXA2 expression is significantly associated with the prognosis of lung cancer patients ( P=6.4e-14). Conclusions:ANXA2 is closely associated with cancer metastasis and holds potential as a new target for metastasis treatment. Further in-depth research will greatly facilitate the transition of ANXA2 from basic research to clinical application.

Objective:To investigate the personality traits of patients with anxious depression and the relationship between personality traits and clinical symptoms.Methods:From December 2011 to October 2014, 177 first-episode untreated patients with depression from the psychiatric department of the First Affiliated Hospital of Kunming Medical University and 185 healthy controls(HC group) recruited by the community were included.All patients were divided into anxious depression group ( n=92) and non-anxious depression group ( n=85) according to whether the anxiety/somatization factor score ≥7.The simplified version of Neuroticism Extraversion Openness Five-Factor Inventory (NEO-FFI) and the Hamilton depression scale-17 (HAMD-17) were used to assess all the subjects.Statistical analyses were conducted in SPSS 21.0.Analysis of covariance was used to compare the differences of the scores on personality dimensions among the three groups.The relationship between personality dimensions and anxious depression was confirmed by Logistic regression, linear regression analysis and generalized linear models. Results:The differences of the scores on the four dimensions of neuroticism ( F=108.863, P<0.01), extraversion ( F=86.357, P<0.01), agreeableness ( F=50.615, P<0.01), and conscientiousness ( F=24.730, P<0.01) among the three groups were statistically significant.Further pairwise comparision showed, the score of neuroticisms was higher in the anxious depression group(43.05±8.92) and non-anxious depression group(39.85±7.21) than that in the HC group (30.16±6.25)( P<0.01, Bonferroni corrected). The scores of extroversion (31.22±6.33, 32.61±6.83), agreeableness (38.66±5.80, 39.46±6.19) and conscientiousness (39.75±6.89, 38.85±7.26) were lower in the anxious depression group and non-anxious depression group than those in the HC group (40.29±5.37, 44.79±4.68, 44.09±5.66, all P<0.01, Bonferroni corrected). The score of neuroticisms in anxious depression group was higher than that in non-anxious depression group, and the difference was statistically significant ( P<0.01, Bonferroni corrected). Logistic regression analysis with age, gender and years of education controlled showed that the score of neuroticism ( B=0.082, OR=1.085, 95% CI=1.020-1.154, P=0.009) and conscientiousness ( B=0.060, OR=1.062, 95% CI=1.006-1.120, P=0.028) were risk factors for anxiety symptoms in patients with depression.Linear regression analysis showed that the scores on neuroticism had positive predictive effects on the anxiety/somatization factor score ( B=0.055, 95% CI=0.021-0.089, P=0.002) and cognitive impairment factor score ( B=0.074, 95% CI=0.023-0.125, P=0.005) in the anxious depression group. Conclusion:Compared to non-anxious depression, patients with anxious depression show higher level of neuroticism, and the level of neuroticism can positively predict the symptoms of anxiety and cognitive impairment.The high level of neuroticism and conscientiousness may be risk factors for the occurrence of anxiety symptoms in patients with depressed.

OBJECTIVE： To investigate the variety and frequency of oral tablets splitting commonly used in pediatrics of children’s hospital， and to explore the feasible scheme of tablet splitting， so as to provide the basis for clinical precision and individuation of drug use. METHODS： From 1st Jul. 2017 to 30th Nov. 2018， all the prescriptions of tablet varieties used for neonates hospitalized in internal and surgical departments in our hospital were obtained. The frequency of each tablet splitting was analyzed， and the literature reports of the tablets splitting were searched. The solubility， stability and recommended method of oral tablets splitting were analyzed， and the suggestion were made. RESULTS： In this study， there were 16 kinds of oral tablets that often needed to be divided into different doses， with a total frequency of 2 149 times. Vitamin B2， Levothyroxine sodium tablets and Ursodeoxycholic acid tablets ranked the top three in terms of dose splitting frequency， accounting for 80.3％ of the total dosing cases. After consulting the literature， the implementation process of the dosage division scheme was formulated， and the splitting methods of commonly used tablets were summarized. It is suggested that clinical evaluation of tablet and patient characteristics should be taken to determine tablet dosing splitting plan. CONCLUSIONS： Tablet dosing splitting is common in neonatal medication in our hospital. The methods of temporary dispensing tablet into oral liquid drugs summarized in this paper can provide references when necessary to meet the needs of clinical individualized drug use.

ngthen cellular immunity, especially Th1-type immune response to HPV11-E7 DNA vaccine in mice.

Objective To obtain the specific IgE antibody related epitopes of Schistosoma japonicum from the phage display library. Methods Serum samples from 150 individials living in the epidemic regions of Schistosomiasis japonica were detected by ABC ELISA. 15 samples with high titer specific IgE antibodies were selected. Their pooled sera were absorbed with Protein G Sepharose beads to remove the IgG antibodies,then,it was used for immunoscreening of a phage display library of random peptide 12 mers. After 5 cycles of screening,DNA samples from 35 phage clones were sequenced. The phage clones with different inserted epitopes were identified immunologically. Results 4 independent phage clones of phage 3,phage 6,phage 8 and phage 15 were determined. Western blotting analysis showed that all of them could be recognized by specific IgE antibodies from the pooled sera. When they were used to immunize BALB/c mice,each clone could cause significant specific IgE antibody response. Conclusions The specific IgE antibody related epitopes of Schistosoma japonicum were screened successfully from the phage display library.

Objective To construct and express human papillomavirus type 11(HPV11) E7 gene with recombinant adenovirus vectors. Methods HPV11 E7 gene was amplified by PCR and directionally cloned into vector pENTR-TOPO to form TOPO-E7 plasmid. E7 gene was transferred into the pAD/CMV/V5-DESTTM gateway vector by LR recombination reaction with pAD/CMV/V5-DESTTM gateway vectors and TOPO-E7 plasmid. The recombination vector was digested by Pac I enzyme and transfected into 293A cell by Lipofectamine method to obtain recombinant adenovirus vectors pAD-E7. Expression of E7 on HaCaT cells infected with pAD-E7 vectors was analyzed by confocal microscopy. Results The recombinant plasmid TOPO-E7 was identified and confirmed with enzyme digestion and sequencing. Recombinant adenovirus vectors pAD-E7 were generated efficiently with a titer of 1.4 ? 107 pfu/mL in transfected 293A cells. E7 protein could be identified in HaCaT cells with confocal microscope 48 h after infected with recombinant adenovirus vector. Conclusions The results indicate efficient expression of HPV11 E7 gene in eukaryotic cells by recombinant adenovirus mediated transfer, which facilitates further research of its function.

A 21-base, Plasmodium falciparum specific, enzyme-conjugated synthetic DNA probe (PFRl-AP) was used for the diagnosis of falciparum malaria. The blood samples (53 P. falciparum, 5 P. vivax, and 3 P. f and P.v mixed infection cases ) collected from Hainan province were tested. The samples of 32 college students were used for normal control. The probe proved to be specific and sensitive. 10pg of purified P.f DNA could be always detected, and there was no cross reaction with the purified DNA of human leukocytes. When testing Hainan blood specimens, PFRl-AP specifically detected P.f infections. In dot blot, when Nytran membrane with 50 microliters of treated blood samples being used, 39 out of 52 P.f specimens hybridized with this probe positively. When the volume of blotted sample was increased to one hundred microliters, the accumulative total positive rate rose up to 88.46%. The samples of P.v and normal control showed negative reaction with this probe.

In this study, the recombinant plasmid pPFl4 labeled with photobiotin was used as a probe to detect the patients infected with Plasmodium falciparum ( P. f.) by dothybridiza-tion. The results showed that out of 35 cases with P. f., 29 were positive, 5 were negative and one was doubtful. One patient with P.f. and P. vivax mixed infection showed positive result. The total positive rate was 83. 3% (30/36). 3 out of 33 normal human blood samples were positive, so the false positive rate was 9%. In addition, there was a correlation between the positive rate of detection and parasitaemia level. The detection sensitivity was 5 ?10-5.