Objective:To assess the performance of machine learning(ML),and integrate the clinical parameters with coronary artery calcium(CAC)score of computed tomography(CT)and quantification of automated epicardial adipose tissue(EAT),so as to predict the long-term risk of myocardial infarction(MI)and cardiogenic death in asymptomatic patients.Methods:A total of 1 058 subjects with cardiovascular risk factors and without symptoms of coronary heart disease who underwent physical examination at the Fifth Medical Center of Chinese PLA General Hospital from January 2013 to October 2015 were selected as this study subjects.A long-term follow-up was conducted on them after CAC score.EAT volume and density were quantified using a fully automated deep learning method.ML extreme gradient boosting was trained by using clinical data,risk score of atherosclerotic cardiovascular disease,CAC score and automated EAT measure,and the repeated 10-fold cross validation was used to verify the model.Results:During the 8-year follow-up period,61 cases of 1 058 subjects occurred events of MI and(or)cardiac death.The area under curve(AUC)value of ML was significantly higher than that of the atherosclerotic cardiovascular disease(ASCVD)risk and the predicting events of CAC score(ML:0.82,ASCVD:0.77,CAC:0.77).Compared with ML with only clinical variable,machine learning based on ASCVD,CAC and EAT had more predictive ability for MI and cardiac death[AUC 0.82(95%CI:77-87)vs.0.78(95%CI:0.72-0.84),P=0.02].The survival rate of subjects with high ML scores had a greater decline degree with the increasing of time,therefore,the subjects with higher ML scores were more likely to experience events.Conclusion:ML,which integrated clinical and quantitative imaging variables,can provide long-term risk prediction for patients with cardiovascular risk factors.

Objective:To summarise the clinical experience in replantation of the severed auricle distal to helix with microsurgical technique.Methods:From December 2018 to October 2021, a total of 5 patients with severed auricle injury were treated in the Department of Hand and Foot Surgery of The Affiliated Hospital of Qingdao University. They were 4 males and 1 female with 23-62 years old. After complete debridement of the auricular pinna, the retrograde replantation method was used. For arteries: a dorsal vein of the foot was used to bridge the posterior auricular artery. For veins: 2 patients had the veins directly anastomosed, 2 had the arteriovenous bridging to the posterior ear vein with dorsal foot veins, and the veins in 1 case were not anastomosed. Among the patients, 2 developed venous occlusions after severed auricle, and were treated with bloodletting through small incision at the skin margin. Two patients who received the arteriovenous of the severed auriclse achieved good blood supply. All the patients underwent treatments including anti-freezing, anti-spasm and anti-infection after the emergency surgery. The follow-ups were conducted regularly by telephone and by display photos via WeChat after surgery.Results:All 5 severed auricles were successfully replanted and survived. Postoperative follow-up ranged from 3 months to 2 years, with an average of 10 months. In the 2 cases with venous crisis, the auricles had mild atrophy. All auricles had no obvious pigmentation, and had the sensation recovery back to normal in 1 year after surgery.Conclusion:The pre-judgment of blood vessel quality and high-quality microsurgery skills are the necessary pre-conditions for auricle replantation. For replantation of severed auricle, it is the key to prevent vascular crisis by having the injured blood vessels thoroughly removed.

[摘 要] 目的：探讨敲减转录因子特异蛋白1（SP1）对小细胞肺癌（SCLC）H466/DDP细胞顺铂（DDP）耐药的影响及其分子机制。方法：构建敲减SP1同时过表达ATP结合盒亚家族C成员1（ABCC1）的SCLC H466/DDP细胞，采用IHC法检测SP1、ABCC1在非耐药和耐药SCLC组织中的表达，用Spearman r法分析SP1与ABCC1在SCLC组织中表达的相关性；WB法检测SP1、ABCC1、CD44在转染后H446/DDP细胞中的表达；CCK-8法、FCM术、微球实验检测转染后H446/DDP细胞的增殖、凋亡及自我复制能力的变化；染色质免疫共沉淀（CHIP）实验检测SP1是否是ABCC1的转录因子。结果：耐药细胞H446/DDP和耐药SCLC组织中的SP1、ABCC1蛋白水平均高于H446细胞和非耐药SCLC组织（均P<0.05），SCLC组织中的SP1、ABCC1蛋白表达呈正相关；敲减SP1抑制H446/DDP细胞的增殖活力，降低CD44、ABCC1蛋白表达水平、减少细胞微球形成数（均P<0.05），促进细胞凋亡（P<0.05）；SP1是ABCC1的转录因子。结论：转录因子SP1通过调控ABBC1的表达影响SCLC H446/DDP细胞的耐药，SP1是SCLC对DDP耐药的潜在治疗靶点。

ObjectiveTo investigate the clinical effect and safety of transcatheter arterial chemoembolization (TACE) combined with microwave ablation (MWA) in the treatment of advanced primary liver cancer. MethodsA total of 186 patients with advanced primary liver cancer who were treated in The Fifth Medical Center of Chinese PLA General Hospital from April 2015 to June 2019 were enrolled and divided into study group and control group using a random number table, with 93 patients in each group. Both groups of patients underwent TACE, and the patients in the study group were treated with ultrasound-guided percutaneous MWA. The two groups were compared in terms of clinical outcome and complications. Quantitative real-time PCR was used to measure the serum level of microRNA-202 (miR-202), ELISA was used to measure the serum levels of fragile histidine triad (FHIT) and P16 protein, and the changes in the above three indices at 3 months after treatment were compared. The two-independent-samples t test was used for comparison of continuous data between two groups, and the paired t-test was used for comparison within one group before and after treatment; The chi-square testwas used for comparison of categorical data between groups. ResultsThe study group had a significantly higher objective response rate than the control group (47.32% vs 27.96%, χ2=7.422, P=0.006), and there was no significant difference in disease control rate between the two groups(P＞0.05). Both groups had significant increases in the serum levels of miR-202, FHIT, and P16 protein at 3 months after treatment (all P＜0.05), and compared with the control group, the study group had significantly higher serum levels of miR-202 (0.84±0.14 vs 0.58±017, t=11.385, P＜0.001), FHIT (1126.35±73.05 pg/ml vs 762.87±56.71 pg/ml, t=37.904, P＜0.001), and P16 protein (52.86±651 pg/ml vs 39.06±5.37 pg/ml, t=15.770, P＜0.001). ConclusionUltrasound-guided MWA in addition to TACE can improve the short-term response of patients with advanced primary liver cancer and increase the serum levels of miR-202, FHIT, and P16 protein, with relatively high safety.

AIM To research the antiasthmatic effects of Zuogui Pills,Yougui Pills and Yupingfeng Powder and their mechanism of action.METHODS Male Brown Norway rats were randomly divided into five groups.OVA was used to induce the model of chronic asthma.Five groups of asthmatic rats were given Zuogui Pills,Yougui Pills,Yupingfeng Powder by gavage once daily for 14 d,respectively.The normal and the model group were given normal saline.Penh value,serum Thl,Th2 inflammatory cytokines,and CROT and ACTH protein levels were determined.Pathological changes of airway remodeling were measured.RT-PCR method was used to measure the expression of CRH mRNA in hypothalamus,and TGF-β1 and Smad3 mRNA in lung tissues.RESULTS Penh values of Yupingfeng Powder group and Yougui Pills group were significantly lower than that of Zuogui Pills group.The Thl inflammatory cytokine of Yupingfeng Powder group was significantly higher than that of Zuogui Pills group,while the Th2 cytokine was significantly lower than that of Zuogui Pills group.In Yougui Pills group,only IL-5 was significantly lower than that of Zuogui Pills group.Serum CROT,ACTH protein and CRH mRNA of Yupingfeng Powder group were significantly higher than those in Zuogui Pills group.No significant difference was found between Yupingfeng Powder group and Zuogui Pills group with regard to TGF-β1 and Smad3 mRNA.Smad3 mRNA of Yougui Pills group was significantly higher than that of the model group.Airway remodeling was significantly reduced by Yupingfeng Powder and Zuogui Pills.The number of goblet cells in Yougui Pills group was significantly lower than that in Zuogui Pills group.CONCLUSION All the three formulaes possess significant antiasthma effects.Regulation effects of Yupingfeng Powder on Thl/Th2 balance,HPA and TGF-β1/Smad3 pathway are strong.Compared with Zuogui Pills,Yougui Pills possesses weaker TGF-β1/Smad3 regulative effects and stronger HPA regulative effects.

BACKGROUND: Neural stem cell transplantation is an emerging therapeutic option in the recovery of neural lesions and neurodegenerative diseases. Neural stem cell culture and differentiation lay a foundation for the further study. OBJECTIVE: To improve the techniques for the isolation, cultivation, differentiation and identification of neural stem cells, and to explore the biological characteristics of cells. METHODS: The neural stem cells from C57BL/6 fetal rats were isolated and cultured in vitro using neurophere culture method followed by morphological and ultrastucture examination. The growth curve and cell cycle of passage 3 cells were drawn and analyzed. Nestin expression was tested by immunofluorescence. Neural stem cells induced in 1% and 10% fetal bovine serum were identified using anti-GFAP, anti-βⅢ-tubulin and anti-MBP by immunofluorescence. RESULTS AND CONCLUSION: The neurospheres exhibited strong cell proliferation ability. Under transmission electron microscope, there was a high nuclear/cytoplasmic ratio in the neural stem cells, indicating a low differentiation degree. Immunofluorescence analysis revealed that neural stem cells were positive for Nestin. The induced cells were positive for GFAP, βⅢ-tubulin, and MBP, indicating these cells were induced to differentiate into astrocytes, neurons and oligodendrocytes, and there were more neurons in 1% fetal bovine serum than those in 10% fetal bovine serum. In conclusion, we could successfully isolate neural stem cells in C57BL/6 mice, and low concentration of fetal bovine serum contributes to more neurons differentiated from neural stem cells.

OBJECTIVE:To systematically review the efficacy of Live combined bacillus subtilis and enterococcus faecium in the prevention and treatment of Antibiotic associated diarrhea(AAD),and provide evidence-based reference for clinical treatment. METHODS:Retrieved from CBM,CJFD and Wanfang database,randomized controlled trials (RCT)of Live combined bacillus subtilis and enterococcus faecium(test group)versus placebo(control group)in the prevention and treatment of AAD. After data ex-tract and quality evaluation,systematical evaluation was performed by using Rev Man 5.0 statistics software. RESULTS:A total of 21 RCT were included,involving 3 881 patients. According to the purpose of different purpose of druguse,the RCT were divided into prevention group and treatment group. Results of Meta-analysis showed the incidence of diarrhea in prevention test group was significantly lower than control group[OR=0.27 ,95% CI(0.22 ,0.32),P<0.001] ,the average hospitalization time in prevention test group was significantly less than control group [MD=-53.19,95%CI(-79.63,-26.75),P<0.001],and the total clinical ef-fective rate in treatment test group was significantly higher than control group[OR=6.76,95%CI(4.16,10.98),P<0.001]. CON-CLUSIONS:The efficacy of Live combined bacillus subtilis and enterococcus faecium are good in the prevention and treatment of AAD. However,due to the limit of methodological quality and sample size,it remains to be further verified with more rigorously designed and long-term follow-up of large-scale RCT.

Objective To explore the clinical application value and feasibility of contrast-enhanced ultrasonography (CEUS) in rapid diagnosis for bleeding of hepatocellular carcinoma. Methods From January 2009 to December 2012, 58 patients from the 302nd Hospital of the People′s Liberation Army underwent CEUS and conventional ultrasound, who were suspicioused primary bleeding of liver cancer or secondary bleeding of liver cancer after percutaneous radiofrequency ablation (RFA). The change of effusion volume in front of liver and in abdominal cavity, the perfusion time, range and characteristics of contrast agent, tumor vessel and the characteristic manifestation of active bleeding within the tumor and subcapsular were evaluated. Theχ2 test was used to compare the show rates of lesion boundary and the bleeding point by conventional ultrasound and CEUS. The t test was used to compare the change of effusion in front of liver and in abdominal cavity from 0 min to 30 min after bleeding were diagnosis by CEUS. Results For 58 cases of suspicious rupture and bleeding of liver cancer, 34 cases were conifrmed by clinical and imaging ifndings. The bleeding points were detected by conventional ultrasound in 5 cases. The bleeding points in 30 cases (including 4 cases of rapid bleeding, 7 cases of medium bleeding and 19 cases of slow bleeding) were detected by CEUS. The detection rate of lesion boundary, tumor vessel and bleeding point of CEUS were higher than those of conventional ultrasound, and the differences were statistically significant (χ2=10.350, P=0.001; χ2=4.300, P=0.034;χ2=36.790, P=0.0007). CEUS showed that contrast agent continuously concentrated at the bleeding point of lesions, and different degrees of bleeding were shown as′jet-like′,′linear′or′intermittent′spillover. The volume of effusion in front of the liver and in abdominal cavity at 30 min was more than those at 0 min after bleeding were diagnosis by CEUS, and the differences were statistically significant (the volume of effusion in abdominal cavity:t=-3.467, P=0.026;t=-12.895, P=0.000;t=-3.055, P=0.007;the volume of effusion in front of the liver:t=-8.110, P=0.001;t=-5.642, P=0.002;t=-5.981, P=0.000). Conclusions CEUS can show direct signs of rupture and bleeding of liver cancer, and the degree of bleeding can be evaluated according to the extravasation characteristics of contrast medium and the changes of lfuid volume. Therefore, CEUS can provid an objective basis for rapid diagnosis of ruptured hepatocellular carcinoma in the emergency scene and bedside.

Objective To study the detection rate of contrast-enhanced ultrasound (CEUS) with different perfusion methods on rabbit VX2 small hepatocellular carcinoma less than 10.0 mm. Methods VX2 tumor cells were inoculated into the subcutaneous tissue of New Zealand rabbit′s thigh. Then the plant tumor were cut into small blocks under sterile conditions and transplanted into hepatic parenchyma in 30 New Zealand rabbits. The contrast media was injected through peripheral vein by single and double perfusion methods. The detection rate of two methods were compared. Results There were 41 hepatocellular carcinoma lesions in the 30 rabbits. There were 15 lesions with size between 3.0 mm and 5.0 mm, and 26 lesions between 5.0 mm and 10.0 mm in diameter. On CEUS, the VX2 tumor presented fast-in and fast-out pattern. In arterial phase, the lesion was enhanced rapidly. In portal venous phase, contrast began to wash out from the carcinoma. In delay phase, the enhancement of lesion was signiifcantly lower than the surrounding normal liver parenchyma. A total of 32 lesions were detected by single perfusion method, including 7 lesions ranging 3.0-5.0 mm and 25 lesions ranging 5.0-10.0 mm. A total of 39 lesions were detected by double perfusion method, including 13 lesions ranging 3.0-5.0 mm and 26 lesions ranging 5.0-10.0 mm. The detection rate of micro-hepatocellular carcinoma by single and double perfusion method was 78% and 95% respectively. The difference was statistically signiifcant (χ2=5.150, P=0.023). The detection rate of 3.0-5.0 mm lesions by single and double perfusion method was 47%and 87%, respectively. The difference was statistically signiifcant ( χ2=5.400, P=0.025). The detection rate of 5.0-10.0 mm lesions by single and double perfusion method was 96% and 100%, respectively. There was no statistically signiifcant difference (χ2=1.020, P=0.500). Conclusion The double perfusion method greatly promotes the detection of micro hepatocellular carcinoma, especially for the lesions less than 5.0 mm in diameter.

Objective To summarize the imaging features of hepatic tumors contained lipid by contrast enhanced ultrasound (CEUS), and compare with MRI ifndings. Methods From July 2010 to December 2012, 17 patients of hepatocellular carcinoma containing lipid component in the 302nd Hospital of the People′s Liberation Army underwent dual-echo sequence MRI imaging. Fifteen patients were conifrmed by enhanced MRI and contrast-enhanced CT, while the other 2 patients were confirmed by pathology. The features of ultrasonography, CEUS and MRI were analyzed. Kappa analysis test was analyzed for the consistency of CEUS and MRI ifndings. Results In the two-dimensional ultrasound of 17 lesions, 9 were hypoechoic heterogeneous lesions, 5 were hypoechoic lesions;13 lesions with clear boundaries, 4 lesions with unclear boundaries;peripheral blood lfow signal in 5 lesions and rich blood lfow in 1 lesion were observed by color Doppler ultrasound, 11 lesions had no signiifcant blood lfow signal;15 lesions showed fast high enhancement in CEUS arterial phase, 1 lesion with synchronization enhancement, 1 lesion with mild enhancement;10 lesions showed portal clearance, 1 lesion showed an enhanced synchronization, 1 lesion with mild enhancement;16 lesions showed clearance in delay phase, 1 enhanced lesion showed isoenhancement;6 lesions presented intra-lesion or surrounding patchy, irregular non-enhancement areas. CEUS diagnostic accuracy was 82%(14/17), the diagnostic accuracy of MRI was 88%(15/17). The Kappa consistency test was 0.628, while CEUS and MRI results were in good agreement. Conclusions Performance of hepatocellular lipid-containing ultrasound contrast arterial phase shows non-enhancement areas. Contrast enhanced ultrasound features of lipid-containing hepatocellular carcinoma are valuable for the clinical diagnois.