Background and purpose:MicroRNA-340 (miR-340) has been demonstrated to play a role of negative regulation in many kinds of tumor, however, there are few reports about the relationship between miR-340 in proliferation and apoptosis of breast cancer cell. This study was aimed to explore the effect of miR-340 on proliferation and apoptosis of breast cancer cell MDA-MB231. Methods: The pre-miR-340 or anti-miR-340 were transiently transfected into breast cancer cell MDA-MB231 with LipofectamineTM2000. miR-340 level was detected by RT-PCR. The Western blot was performed to detect the protein level of cleaved-caspase-3. The inhibition rate of cell proliferation was evaluated by MTT assay. The cell apoptosis was studied by lfow cytometry. Results:The pre-miR-340 facilitated the expression of miR-340 in MDA-MB231 cells. The pre-miR-340 enhanced the protein level of cleaved-caspase-3, inhibited the proliferation of MDA-MB231 cells and increased its apoptosis. On the contrary, the expression of miR-340 was inhibited by anti-miR-340 in MDA-MB231 cells. The protein level of cleaved-caspase-3 was reduced after the anti-miR-340-transfected MDA-MB231 cells. Anti-miR-340 promoted the proliferation of MDA-MB231 cells. Decreased apoptosis of MDA-MB231 cells was observed by lfow cytometry. Conclusion:The overexpression of miR-340 can effectively inhibit the proliferation and increase the apoptosis of MDA-MB231 cells, which may be explained by up-regulating of protein cleaved-caspase-3 level.

A robust,selective and highly sensitive chemiluminescent (CL) platform for protein assay was presented in this paper.This novel CL approach utilized rolling circle amplification (RCA) as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier.Typically,the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence.This aptamer-primer sequence was then employed as the primer of RCA.Based on this design,a number of the biotinylated probes and streptavidin-horseradish peroxidase (SA-HRP) were captured on the plate,and the CL signal was amplified.In summary,our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized,and devoid of light source.Therefore,this new technique will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases,by taking advantages of high sensitivity and selectivity.

A robust, selective and highly sensitive chemiluminescent （CL） platform for protein assay was presented in this paper. This novel CL approach utilized rolling circle amplification （RCA） as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier. Typically, the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence. This aptamer-primer sequence was then employed as the primer of RCA. Based on this design, a number of the biotinylated probes and streptavidin-horseradish peroxidase （SA-HRP） were captured on the plate, and the CL signal was amplified. In summary, our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized, and devoid of light source. Therefore, this new technique .will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases, by taking advantages of high sensitivity and selectivity.

Objective:To explore the changes of number of CD4+ CD25 + foxp3 + regulatory T cells (Treg)and natural killer cells (NK)in the peripheral immune organs and tumor tissue of the murine models of lung cancer,and to clarify their effects on the development of lung cancer.Methods:The C57BL/6 mice were divided into Lewis lung carcinoma cells (LLC)injection group and normal control group.The mice in LLC injection group were injected with LLC subcutaneously in the armpit to establish the tumor models,while the mice in normal control group were injected with the same amount of saline.The number of CD4+ CD25 + T cells,CD4+ CD25 + foxp3 + Tregs in the spleen,lymph nodes and lung cancer tissues,and the number of NK cells in the spleen tissue were labeled by cell surface or intracellular antibody staining,and detected by flow cytometry.Results:The ratios of CD4+ CD25 + T cells to CD4+ T cells,foxp3+ cells to CD4+ CD25 + T cells,and the number of CD4+ CD25 + foxp3 + Treg in the spleen and lymph nodes of the mice in LLC injection group were increased significantly compared with normal control group (P <0.05 or P <0.01).Moreover,the ratios of CD4+ CD25 + T cells to CD4+ T cells and foxp3 + cells to CD4+ CD25 + T cells in the tumor tissue were significantly higher than those in the spleen and lymph nodes of the mice in LLC injection group.However,the ratio of NK cells in the spleen tissue of the mice in lung cancer group was significantly decreased compared with normal control group (P <0.05).Conclusion:The increase of ratio and the number of Treg cells and the decrease of ratio of NK cells in the main immune organs of lung cancer mice may promote the development of tumor and inhibit the immune response to cancer cells in vivo .

Objective:To investigate the change of nature killer T cell(NKT)and myeloid derived suppressor cells(MDSC) during the development of mouse lung tumor.Methods:Lung tumor mouse models were made by subcutaneous injection of Lewis lung tumor cells( LLC) ,peripheral blood leukocytes were extracted from mouse tail blood at different time points after LLC injection.NKT and MDSC were detected by flow cytometry after relative antibody staining.Results:With the increasing volume of lung tumor,the ratio of NKT cells decreased gradually,while the ratio of MDSC increased gradually in the peripheral blood of LLC-injected mice.Both NKT and MDSC showed significantly changes in LLC-injected mice compared with that of normal control mice.Conclusion:NKT and MDSC in LLC-injected mice show opposite changes during the development of lung tumor,so,they can be used as potential monitoring index for lung tumor development.

Objective To observe the missed diagnosis of invasive carcinoma under the microscope (ICUM) in high grade squamous intraepithelial neoplasia (HSIL), and analyze associated factors influencing missed ICUM. Methods A retrospective study was performed on patients diagnosed with HSIL by colposcopy-guided biopsy and treated with loop electrosurgical excision procedure(LEEP)at the First Affiliated Hospital of Nanjing Medical University, from December 2014 to December 2016. They were non-pregnant, ≤50 years old and the cervical volume without obvious enlargement and exogenous surface without and ulcerative lesions. A total of 283 cases with early cervical cytology results, never received cervical traumatic treatment or cervical biopsy in another hospital before, and their colposcopic images were clear enough to reevaluate. The ultimate pathological diagnosis was based on the higher-level pathological diagnosis between the results of cervical biopsy and LEEP to evaluate ICUM missed in HSIL and the risk factors. Results (1) Among the 283 cases with HSIL diagnosed by colposcopy-directed biopsy,44 cases (15.5％,44/283) were missed diagnosis of ICUM, which consisted of 29 cases Ⅰa1, 4 cases Ⅰa2 and 11 cases Ⅰb1 in the ultimate pathology.(2)Analysis of associated factors for missed ICUM:univariate analysis showed that,as the age increased, the risk of missed ICUM also increased(the rates of missed diagnosis for<30, 30-39, 40-50 years were 7.7％, 11.5％, 22.0％, respectively;χ2=6.254, P=0.012 by trend test). The more the number of high-grade features, the higher risks(the rates of missed diagnosis for 1, 2, 3, 4 high-grade features were 10.2％, 17.6％, 23.8％, 30.8％, respectively;χ2=7.686,P=0.006 by trend test). The locations of HSIL were only endocervical, only ectocervical and mixed, the risk increased by this sequence(2.8％, 5.1％, 28.7％; χ2=26.193,P<0.01 by trend test). The rate of missed diagnosis for not completely visible squamocolumnar junction(SCJ)was higher than that of the completely visible one(22.3％ vs 2.1％;χ2=19.680, P<0.01). The rate of missed diagnosis was higher for existing atypical vessels than those without(60.7％ vs 10.6％;χ2=48.279, P<0.01). The rate of missed diagnosis for visible lesion size≥40 mm2 was higher than that of<40 mm2(27.3％vs 4.2％;χ2=28.921, P<0.01). The rate of missed diagnosis for the proportion of visible lesion size in ectocervical size ≥0.75 was higher than that of <0.75 (83.3％ vs 14.1％;P<0.01). The rate of missed diagnosis for the maximum linear length of visible lesion≥10 mm was higher than that of<10 mm(46.9％vs 9.0％;χ2=44.473, P<0.01). But the different severity of cervical cytology before colposcopy was not associated with missed ICUM(P>0.05). Multivariable analysis found that visibility of SCJ, atypical vessels, visible lesion size and maximum linear length of visible lesion were associated with missed diagnosis of ICUM(all P<0.05). Conclusions The diagnostic value of HSIL by colposcopy is limited. Meanwhile, for the patients who are ≤50 years old with HSIL diagnosed by cervical biopsy, invisibility of SCJ, atypical vessels, visible lesion size and maximum linear length of visible lesion evaluated by colposcopy are the independent risk factors of missed ICUM. Thereby, it is necessary to take active intervention for HSIL with these risk factors.

Objective Toexplorethevalueofmulti-parameterquantitativeanalysisofspectralCTimaging (GSI)indifferentiatingrenal fat-poorangiomyolipoma(fpAML)andchromophobecellrenalcarcinoma(CCRC).Methods 42patientswithrenaltumor,including 25caseswithfpAMLand17caseswithCCRC,wereretrospectivelyanalyzed.Allofthem werescannedinGSImode.Themorphology differencesbetweenthefpAMLgroupandtheCCRCgroupwereanalyzed.GSIViewersoftwarewasusedtocalculatetheiodineconcentration (IC),thenormalizediodineconcentration(NIC),thesloperateofthespectrumenergycurveinthecorticalphase(CP)andmedullaryphase (MP),respectively.Thedifferencesofthoseparameterswerecomparedbetweenthetwogroupsusingthetwo-simplettest.Results Somecharacteristicsigns,suchas"blackspots"sign,cracksignandnecrosishadthevaluefordifferentialdiagnosis.IntheCP,theIC ofthefpAMLandCCRCgroupwere30.20±5.25vs19.97±4.01,theNICswere0.45±0.10vs0.32±0.06,andthesloperatesof spectrumenergycurveswere3.45±1.23vs2.42±0.48,respectively.IntheNP,theICofthefpAMLandCCRCgroupwere27.84± 8.07vs22.94±4.46,theNICswere0.58±0.17vs0.46±0.11,andthesloperatesofthespectrumenergycurveswere3.24±1.25vs 2.69±0.47,respectively.Thereweresignificantdifferencesbetween2groups(P<0.05).TheNICintheCPprovidedhighsensitivity (75%)andspecificity(86%)indifferentiatingfpAMLfrom CCRC,andtheareaundertheROCcurvewas0.886.Conclusion The focalcysticandnecrotic,enhanceduniformityanddegree,"blackspots"sign,cracksignand multi-parametersofGSI,includingIC, NIC,andthesloperateofthespectrumenergycurvecouldplayimportantroleindifferentialdiagnosisbetweenfpAMLandCCRC.

Objective To analyze the performance of colposcopy and investigate the diagnosis and treatment characteristics of high-grade squamous intraepithelial lesion (HSIL) diagnosed by cervical tissue sampling in post-menopausal women. Methods A retrospective study was performed on 1 449 patients with HSIL diagnosed by cervical tissue sampling under colposcopy and treated by loop electrosurgical excision procedure (LEEP) or extrafascial hysterectomy as the primary therapy at the First Affiliated Hospital of Nanjing Medical University, from November 2015 to October 2017. In order to investigate the diagnosis and treatment of HSIL in post-menopausal women, a case-control study was conducted to compare the difference in performance of colposcopy and treatment modality between 213 post-menopausal patients (14.7%, 213/1 449) and 1 236 pre-menopausal patients (85.3%, 1 236/1 449). Results (1)The proportion of cases pathologically upgraded to cervical cancer was significantly greater in post-menopausal patients (9.4%, 20/213) compared with pre-menopausal patients (3.8%, 47/1 236; P<0.05). (2) The proportion of ≥HSIL diagnosed by colposcopy showed no significant difference between post-menopausal patients (76.1%, 162/213) and pre-menopausal patients (78.2%, 967/1 236; P=0.479). The proportion of typeⅢtransformation zone (TZ) was significantly greater in post-menopausal patients (91.1%, 194/213) compared with pre-menopausal patients (59.1%, 731/1 236; P<0.05). The rate of missed diagnosis of cervical cancer was significantly higher in typeⅢTZ (6.4%, 59/925) compared with typeⅠand(or)ⅡTZ (1.5%, 8/524; P<0.05). The proportion of HSIL detected by endocervical curettage alone was greater in post-menopausal patients (9.9%, 21/213) compared with pre-menopausal patients (2.6%, 32/1 236; P<0.05). (3)Initial treatment with LEEP: the positive rate of endocervical margin was significantly greater in post-menopausal patients (20.5%, 36/176) compared with pre-menopausal patients (10.5%, 130/1 236;P<0.05); in patients who were diagnosed as HSIL after LEEP, the positive rate of endocervical margin and the residual rate were both greater in post-menopausal patients compared with pre-menopausal patients [15.4% (25/162) versus 8.8% (105/1 189), P=0.008; 52.0% (13/25) versus 26.7% (28/105), P=0.014]. (4)Thirty-seven post-menopausal patients were treated by extrafascial hysterectomy as the primary therapy, 5 cases (13.5%, 5/37) were diagnosed as cervical cancer (stage Ⅰa1) after the surgery. Conclusions (1) The lesions of HSIL in post-menopausal patients still have definite features under colposcopy as same as pre-menopausal patients. Endocervical curettage could help detect more HSIL in post-menopausal patients. Compared with pre-menopausal patients, post-menopausal HSIL patients have an increased risk of cervical cancer and are more likely missed by cervical tissue sampling. (2) LEEP has the dual effects of diagnosis and treatment, and is still the recommended treatment for post-menopausal HSIL patients. However, the increase in positive rate of endocervical margin and residual rate requires further active intervention. (3) Considering those post-menopausal HSIL patients who cannot accept conization as the initial treatment, the selection of hysterectomy type requires more thorough study.

Objective:To explore the effect of virtual reality (VR) technology in postoperative pain management for surgical patients.Methods:The literature on VR technology for relieving postoperative pain in surgical patients was electronically retrieved from eight databases, including PubMed, Web of Science and so on. The search period was from database establishment to November 19, 2023. Two researchers independently screened literature, extracted data, and conducted literature quality evaluation, using RevMan 5.4 for Meta-analysis.Results:A total of 18 articles were included. Meta-analysis results showed that VR technology could reduce postoperative resting pain scores [ MD=-1.13, 95% CI (-1.29, -0.96), P<0.001], postoperative 12 hour pain scores [ MD=-0.49, 95% CI (-0.73, -0.24), P<0.001], postoperative 24 hour pain scores [ MD=-0.73, 95% CI (-0.92, -0.54), P<0.001], postoperative 48 hour pain scores [ MD=-0.69, 95% CI (-0.84, -0.53), P<0.001], postoperative 72 hour pain scores [ MD=-0.37, 95% CI (-0.59, -0.16), P<0.001], and postoperative pain scores during dressing changes [ MD=-1.11, 95% CI (-2.06, -0.16), P=0.02], and could improve patient postoperative comfort [ MD=8.31, 95% CI (6.57, 10.06), P<0.001] . Conclusions:As a non-pharmacological intervention, VR technology can alleviate postoperative pain and improve patient comfort in surgical patients. In the future, large-scale and high-quality research is still needed to further verify the effectiveness of VR technology.

Objective:To explore the current situation and influencing factors of amputation decision-making dilemma of diabetic foot patients.Methods:From July to December 2023, 200 patients with diabetic foot in the Tianjin Medical University General Hospital and Tianjin Medical University Chu Hsien-I Memorial Hospital were selected as study subjects by convenience sampling. General Information Questionnaire, Decisional Conflict Scale (DCS), Family APGAR Index, and Hospital Anxiety and Depression Scale were used to conduct a cross-sectional survey. Pearson correlation was used to analyze the correlation between diabetic foot patients' amputation decision-making dilemma and family caring, anxiety and depression, and multiple linear regression was used to analyze the influencing factors of diabetic foot patients' amputation decision-making dilemma.Results:A total of 200 questionnaires were distributed, and 180 valid questionnaires were collected, with a valid response rate of 90.0% (180/200). The DCS score of 180 patients with diabetic foot was (30.04±9.77), 76.7% (138/180) patients scored ≥25.0, and they had decision-making dilemma, and 25.0% (45/180) of patients scored ≥37.5, indicating decision-making delay. Multiple linear regression analysis showed that occupational status, diabetes course, family caring, anxiety and depression were the influencing factors of amputation decision-making dilemma of diabetic foot patients ( P<0.05) . Conclusions:Diabetic foot patients face certain dilemmas in the process of amputation decision-making. Clinical medical and nursing staff should reasonably evaluate the patient's occupational status, disease course, family caring, and psychological state, and develop personalized decision support strategies to improve decision quality and prevent changes in the patient's condition caused by delayed decision-making.