Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter.
Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively.
Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.

Objective To observe the blood flow changes of left ventricular cavity and quantify the energy loss (EL) and circulation of left ventricular during systole and diastole in patients with dilated cardiomyopathy(DCM) via vector flow mapping(VFM).Methods Thirty-six healthy volunteers and 32DCM patients were enrolled.According to the severity of mitral regurgitation(MR),DCM patients were divided into two groups.The quantitative parameters,including average energy loss(EL-base,EL mid,EL-apex) and circulation (vortex quantity,vortex area,circulation) were measured in the different periods of VFM imaging mode in apical four-chamber view,apical three-chamber view and apical two-chamber view respectively.The difference of parameters was evaluated between different groups during the different periods.The early transmitral valve blood flow velocity E,the late transmitral valve blood flow velocity A,the ratio of early transmitral valve blood flow velocity to the early diastolic mitral valve velocity E/e,the ratio of early transmitral valve blood flow velocity to the late transmitral valve blood flow velocity E/A and Tei index were derived via dual-Doppler imaging technology.Meanwhile,the correlation between VFM parameters with E,A,E/A,E/e or Tei index was also analyzed.Results ①The level of EL were decreased significantly among EL-mid and EL-apex during early diastole,EL-mid during mid-systole,EL-mid and EL-apex during late-systole (P ＜0.05),when compared with the control group.The vortex quantity,vortex area and circulation during early diastole and the vortex area and circulation during atrial contraction period were significantly different between two groups (all P ＜ 0.05).As for mid-diastole,only vortex area showed significant difference (P ＜0).01).(②) When compared with different grades of MR in DCM patients,the level of all segments EL during diastole and EL-base during systole increased with severity of MR(P ＜0.05).The circulation during diastole also increased with the severity of MR(P ＜0.05).③The vortex area and circulation had positive correlation with E/e during diastole in all subjects(all P ＜0.05).The vortex area during atrial contraction period and early diastole had positive correlation with E/A and Tei index in all subjects(r =0.630,0.345,0.468,0.316,0.663,0.264,respectively,allP ＜0.01).EL-mid during late-systole were negatively correlated with LV end diastolic volume,end systolic volume,but positively correlated with ejection fraction in all subjects(r =0.335,-0.312,0.340,respectively,all P ＜0.05).While,there was positive correlation between E/A with the basal segment of mid-diastole(r =0.376,P ＜0.01).Conclusions The visualization of hydromechanics inside the heart of DCM patients could be directly accomplished by VFM,which can provide a new method for diagnosis and evaluation of DCM.

Livin,as a novel member of human inhibitor of apoptosis protein family,is highly expressed in many malignant tumors.Livin plays critical role in apoptosis inhibition,regulating the cell cycle,participating in tumor angiogenesis.Livin is also significant to chemoresistance.The majority of the current data suggests that Livin expression in cancer appears to be associated with unfavorable clinico-pathological parameters,such as disease relapse and shorter patient survival.In recent years,immunotherapy and gene therapy for the targeting of Livin have become a hot research field,which provides new strategy and direction for tumor therapy.

Objective To study the expressions of survivin and PTEN in ampullary carcinoma and their clinical significance.Methods The expressions of Survivin and PTEN proteins were tested by EnVision immunohistochemistry in 40 cases of ampullary carcinomas and 8 cases of normal ampulla of vater as control.Results The positive rate of Survivin in ampullary carcinomas was significantly higher than normal human controls (82.5％ vs 0％,P ＜ 0.01 ).The differences of the expression of Survivin in ampullaty carcinoma of duodenal invasion,pancreatic invasion,and lymph node metastasis were significant ( P ＜0.05 ).Significant differences were also observed in the expression rate of PTEN hetween the patients with ampullary carcinoma and the normal controls (50％ vs 100％,P ＜ 0.01 ).The differences of the expression of PTEN in ampullary carcinoma of duodenal invasion,pancreatic invasion,and lymph node metastasis were significant ( P ＜ 0.05 ).Significantly negative correlation was found between the expression of Survivin and PTEN by using spearman correlation analysis ( r =-0.57,P ＜ 0.01 ).Conclusions The abnormal expression of Survivin and PTEN gene may promote tumor genesis and progression of ampullary carcinomas and it may be used as the marker for prognosis.

Objective To investigate the clinical efficacy of SOX and XELOX in neoadjuvant chemo-therapy for advanced gastric carcinoma .Methods Seventy-five cases with advanced gastric carcinoma were se-lected from our hospital from Feb 1,2011 to Oct 1,2015.SOX and XELOX were used to these patients treatment . The relationship between SOX and XELOX with tumor stage ,clinical curative effect evaluation ,operation ,postop-erative pathology ,adverse reaction ,survival analysis with advanced gastric carcinoma had been analyzed retrospec -tively.Results The efficacy of SOX and XELOX were 47.22%,41.03%.Compared with the two groups ,the CT curative effect evaluation,effective rate,disease control rate had no significant differences (P>0.05).The median survival time of DFS and OS were significant between the two groups (P<0.05).Conclusion The SOX and XELOX for neoadjuvant chemotherapy of advanced gastric carcinoma treatment is safety and effective , and the XELOX is better than SOX in terms of long -term effect,prognosis and clinical benefits .

ObjectiveTo study the expressions of Survivin and Bcl-2 in carcinoma of the ampulla of Vater and their clinical significance.Methods The expressions of Survivin and Bcl-2 proteins were tested by EnVision immunohistochemistry in 40 cases of ampullary carcinomas,and 8 cases of normal ampulla of rater as control.ResultsThe positive expression of Survivin and Bcl-2 in the tissues from the ampullary carcinoma was significantly higher than that in the normal controls(82.5％,72.5％ vs 0％,12.5％,P ＜0.05).The expression of Survivin and Bcl-2 had no relations with the sex,age,tumor size,histological types,and clinical stages( P ＞ 0.05 ).The differences of the expression of Survivin and Bcl-2 in ampullary carcinoma of duodenal invasion,pancreatic invasion,and lymph node metastasis were significant(P ＜ 0.05) ; Significantly positive correlation was found between the expression of Survivin and Bcl-2 by using spearman correlation analysis( r =0.647,P ＜ 0.05 ).Conclusions Survivin and Bcl-2 may play an important role in tumorigenesis of ampullary carcinoma.Combined detection of Survivin and Bcl-2 may be useful to determine the degree of malignancy and the progress of the ampullary carcinoma.

Objective:To explore the present situation of medical students' professionalism cognition and provide reference for medical personnel training and medical education.Methods:A cross-sectional survey was conducted on medical students among 2 affiliated hospitals of a medical university in H province by cluster sampling. Exploratory factor analysis and reliability and validity test were carried out and a descriptive analysis of the present situation of professionalism was conducted by SPSS 20.0. Amos21.0 software was used for a confirmatory factor analysis on the samples.Results:The measurement scale had good reliability and validity, and the Cronbach alpha coefficient of the scale was 0.901. The average professionalism score of the medical students was about 80 points. The score of "physical and mental status and self-development ability" was the highest (83.65 points), and the lowest score was "academic ability" (72.21 points). There was a little difference in the professionalism cognition between the two hospitals, with significant differences between the dimension "respect and care" and "responsibility".Conclusion:This study has initially formed a medical professionalism measurement scale with good reliability and validity. The professionalism of medical students in the 2 affiliated hospitals of a medical school in Province H is in good condition as a whole. In the future, medical education should pay more attention to the combination of basic professional knowledge and clinical practice of medical students, and change the training model of medical students in order to improve the academic ability of medical students and medical students' overall cognition of professionalism.

The efficient clinical treatment of oral squamous cell carcinoma(OSCC)is still a challenge that demands the development of effective new drugs.Phenformin has been shown to produce more potent anti-tumor activities than metformin on different tumors,however,not much is known about the influence of phenformin on OSCC cells.We found that phenformin suppresses OSCC cell proliferation,and promotes OSCC cell autophagy and apoptosis to significantly inhibit OSCC cell growth both in vivo and in vitro.RNA-seq analysis revealed that autophagy pathways were the main targets of phenformin and identified two new targets DDIT4(DNA damage inducible transcript 4)and NIBAN1(niban apoptosis regulator 1).We found that phenformin significantly induces the expression of both DDIT4 and NIBAN1 to promote OSCC autophagy.Further,the enhanced expression of DDIT4 and NIBAN1 elicited by phenformin was not blocked by the knockdown of AMPK but was suppressed by the knockdown of transcription factor ATF4(activation transcription factor 4),which was induced by phenformin treatment in OSCC cells.Mechanistically,these results revealed that phenformin triggers endoplasmic reticulum(ER)stress to activate PERK(protein kinase R-like ER kinase),which phosphorylates the transitional initial factor eIF2,and the increased phosphorylation of eIF2 leads to the increased translation of ATF4.In summary,we discovered that phenformin induces its new targets DDIT4 and especially NIBAN1 to promote autophagic and apoptotic cell death to suppress OSCC cell growth.Our study supports the potential clinical utility of phenformin for OSCC treatment in the future.

OBJECTIVE: To explore the strategies of prenatal diagnosis and genetic counseling for fetuses of two families with large deletions of 13q21. METHODS: Two singleton fetuses who were diagnosed with chromosome 13 microdeletions by non-invasive prenatal testing (NIPT) at Ningbo Women and Children's Hospital in March 2021 and December 2021 respectively were selected as the study subjects. Chromosomal karyotyping and chromosomal microarray analysis (CMA) were carried on amniotic samples. Peripheral blood samples were collected from the two couples for CMA assay to determine the origin of abnormal chromosomes identified in the fetuses. RESULTS: The karyotypes of the two fetuses were both normal. CMA revealed that they have respectively harbored heterozygous deletions spanning 11.935 Mb at 13q21.1q21.33 and 10.995 Mb at 13q14.3q21.32, which were respectively inherited from their mother and father. Both deletions had low gene density and lacked haploinsufficient genes, and were predicted to be likely benign variants based on database and literature search. Both couples had opted to continue with the pregnancy. CONCLUSION The deletions of the 13q21 region in both families may be of benign variants. As the follow-up time was short, there was no sufficient evidence for the determination of pathogenicity, though our finding may still provide a basis for the prenatal diagnosis and genetic counseling.
Pregnancy ; Child ; Female ; Humans ; Pedigree ; East Asian People ; Prenatal Diagnosis ; Chromosome Aberrations ; Karyotyping ; Microarray Analysis ; DNA Copy Number Variations

Objective:This study analyzes COVID-19 prevention and treatment technology patents from 103 countries and regions worldwide, in order to have a good grasp of the most-current trends in COVID-19 prevention and treatment technology research worldwide, discuss the research directions of such technologies, provide theoretical foundations for interdisciplinary and multi-team cooperation.Methods:The retrieval strategies were constructed utilizing the retrieval and analysis system by the Regional Patent Information Service (Nanjing) Center of The State Intellectual Property Office and IncoPat's scientific and technological innovation platform, using third-level branches as the unit and separating the retrieval and proof processes, the queries built using IPC classification numbers and key words; the search was limited to invention patents, with utility models and designs excluded.Results:On February 14, 2020, there had been a total number of 136 087 invention patent applications for COVID-19 prevention and treatment technologies globally. In terms of the country of origin, the U. S. ranked first with 56 095 patents, followed by China (18 096). In terms of technical field, the top three were chemical therapeutic agents (47 634), vaccines (47 248) and diagnostic tests (12 991), with the U. S. possessing absolutely advantageous patent portfolios in all three fields. The applicants were mainly comprised of large enterprises abroad and universities and research institutes in China.Conclusions:China′s COVID-19 related patent applications are relatively scattered. In the field of chemotherapy, there is room for improvement compared with the advanced countries, and cooperation between scientific institutions and enterprises is insufficient. Therefore, China should further increase investment in chemical drug research and development and seize the opportunity to actively promote the research and development of vaccine technology. On the other hand, we need to consolidate the technical advantages and product advantages of epidemic prevention and control which are already accumulated, and strengthen the protection and layout of intellectual property rights. Furthermore, the technological innovation system combining production, education and research is the breakthrough point to encourage enterprises to become the subject of technological innovation.Last but not least, we should build a service platform for the transfer and transformation of scientific and technological achievements to enhance intellectual property rights and promote the transformation of scientific and technological achievements.