Objective:To establish a long-term mortality rate prediction model for patients aged 60 years and over with atrial fibrillation and coronary heart disease using the machine learning method, and identify the corresponding risk factors of mortality.Methods:In this retrospective cohort study, a total of 329(11 cases lost of follow-up)patients with 183 males(55.6%)and 146 females(44.4%), aged(77.8±7.3)years, and 142 patients aged 80 years or older(43.2%)were selected in our hospitals from January 2013 to March 2015.And their clinical data on atrial fibrillation and coronary heart disease were analyzed.They were divided into the death group(151 cases)and the survival group(167 cases)according to the survival outcome.In addition, 60 patients aged 60 years and over admitted to our hospitals from April to July 2015 with atrial fibrillation and coronary heart disease were selected as external data validation set.The clinical data included age, gender, body mass index, diagnosis, co-morbidity, laboratory indicators, electrocardiogram, echocardiogram, treatment data.These patients were followed up for at least 6 years, and the main adverse cardiovascular and cerebrovascular events(MACCE), including death, were recorded.Finally, the data of the enrolled patients were randomly divided into the training set and the test set according to the ratio of 9∶1, Different models were established to predict the long-term mortality of patients with atrial fibrillation and coronary heart disease by machine learning algorithm.The optimal model was established by substituting external data(60 cases)into the model for verification and comparison.The top 20 risk factors for mortality were determined by Shapley additive explanation(SHAP)algorithm.Results:A total of 329 hospitalized patients were included in this study, the overall median follow-up time was 77.0 months(95% CI: 54.0~84.0), 11 cases lost during follow-up(3.3%), and 151 cases died(45.9%). The analysis found that the areas under the ROC curve for a support vector machine(SVM)model, k-Nearest Neighbor(KNN)model, decision tree model, random forest model, ADABoost model, XGBoost model and logistic regression model were 0.76, 0.75, 0.75, 0.91, 0.86, 0.85 and 0.81, respectively.The random forest model had the highest prediction efficiency, with the accuracy of 0.789 and F1 value of 0.806, which was better than the logistic regression model[the Area Under Receiver Operating Characteristic Curve(AUC): 0.91 vs.0.81, P<0.05]. D-dimer, age, number of MACCE, left ventricular ejection fraction, serum albumin level, anemia, New York Heart Association(NYHA)grade, history of old myocardial infarction, estimated glomerular filtration rate(eGFR)and resting heart rate were important risk factors for predicting long-term mortality. Conclusions:The random forest model based on machine learning method can predict the long-term mortality of patients with atrial fibrillation and coronary heart disease aged 60 years and over, have a good identification ability.Its accuracy is higher than that of the traditional Logistic regression model.Reducing the long-term mortality and improving the long-term outcomes can be achieved by intervening on D-dimer levels, correcting hypoproteinemia and anemia, improving cardiac function and controlling resting ventricular rates.

OBJECTIVETo explore the genetic etiology for 11 sporadic patients with neurofibromatosis type 1.

METHODSChip targeting capture and high-throughput sequencing were employed to detect potential mutations of NF1 and NF2 genes among the 11 patients. The data was filtered through multiple mutational databases and in-house whole exome sequence database. Sanger sequencing was used for analysis of family members of the patients.

RESULTSEleven pathogenic variants were found among the 11 patients, which included two splicing mutations, one missense mutation, two nonsense mutations, and six frame-shifting mutations. None of the mutations was recorded by the public database or the in-house database generated from 1775 samples through whole exome sequencing. None of the unaffected parents carried the same mutation. Seven mutations were associated with neurofibromatosis type 1 previously, while the remaining four were discovered for the first time. Prenatal diagnosis of two high-risk pregnancies suggested that neither fetus has inherited the NF1 mutation from their affected parents.

CONCLUSIONIdentification of causative mutations in patients with sporadic-type neurofibromatosis type 1 has provided a basis for genetic counseling. The four novel mutations have enriched the spectrum of NF1 gene mutations.

Objective To prepare 99 Tcm-HYNIC-c( isoDGRKy) as a SPECT/CT imaging molecu-lar probe targeting integrin αvβ3 , and evaluate its biodistribution and feasibility on SPECT/CT imaging for integrinαvβ3-positive tumor in U87MG human glioma xenograft mouse models. Methods The bifunctional chelator HYNIC was conjugated to c( isoDGRKy) , and tricine and TPPTS were used as coligands for 99 Tcm labeling to prepare 99 Tcm-HYNIC-c( isoDGRKy) . The radiochemical purity and stability of the product were measured. The expression of integrin αvβ3 and binding affinity ( half maximal inhibitory concentration, IC50 ) of c ( isoDGRKy ) was detected in U87MG cells by cell experiments in vitro. Biodistribution and SPECT/CT imaging of 99 Tcm-HYNIC-c( isoDGRKy) including blocking experiments were performed respec-tively in nude mice bearing U87MG human glioma xenografts. Results The radiochemical purity of 99 Tcm-HYNIC-c( isoDGRKy) was over 99%, and was still over 99% after 4 h incubation in saline at room temper-ature. Flow cytometry assay showed that U87MG cells were integrinαvβ3-positive ( expressive rate:70%) . The IC50 of c(isoDGRKy) was 6.67×10-8 mol/L. Biodistribution results showed 99Tcm-HYNIC-c(isoDGRKy) with a rapid clearance from blood was excreted mainly via the kidneys. The 99 Tcm-HYNIC-c( isoDGRKy) uptake values in U87MG tumors were (7.31±1.42) and (1.09±0.11) %ID/g at 15 and 45 min post-injection re-spectively, and tumor-to-muscle ratio reached 5.01±1.47 at 15 min post-injection. The tumors were clearlyvisualized with low background from 0.5 to 1 h post-injection in tumor bearing mice. In the blocking experi-ment, the tumor was barely visualized after co-injection of excess cold c(RGDfK) peptide with 99Tcm-HYNIC-c(isoDGRKy). Conclusions 99Tcm-HYNIC-c(isoDGRKy) may be easily and steadily prepared. It may be a RGD-like promising SPECT/CT imaging probe for integrinαvβ3-positive tumor.

Objective To prepare a novel dual-modality imaging probe based on Cerasome nano-materials, and evaluate its in vivo biodistribution and pharmacokinetic properties. Methods ICG encapsu-lated Cerasome was modified with chelating agent DOTA for 111 In-labeling. Normal mice firstly were used for in vivo studies. Animals were sacrificed at different time points after tail vein administration, blood samples were taken and the organs of interest were captured to evaluate the pharmacokinetic properties and in vivo biodistribution of 111 In-ICG-DPDCs. The subcutaneous Lewis lung carcinoma ( LLC ) tumor model in C57BL/6 mouse was established. The tumor-bearing mice were subjected to optical imaging in small animal IVIS and SPECT imaging in small animal nanoScanSPECT/CT system for tumor uptake of 111 In-ICG-DPDCs. Results The size of the nanoparticle probe was about 90 nm, and the 111 In-labeling was successfully per-formed with 99.93% radiochemical purity after purification. 111 In-ICG-DPDCs showed excellent in vitro sta-bility with 97.10% radiochemical purity at 48 h post-purification. In vivo blood clearance experiments showed that 111 In-ICG-DPDCs had a relative long blood circulation time with the fast and slow phase half-lives of 40 and 132.7 min. 111In-ICG-DPDCs accumulated mainly in the liver and spleen, with long retention time. NanoScanSPECT/CT imaging showed that LLC tumors were significantly visualized at 4 h post-injection, and the other major accumulated organs were the liver and spleen, which were consistent with the results of biodistribution. Optical imaging showed significant uptake of the nanoparticle probe in the tumor, confirming the SPECT imaging results. Conclusion The Cerasome based probe designed could be used for tumor SPECT and optical dual-modality imaging, and has potential for therapeutic use.

Objective To investigate the clinical application of temporary balloon occlusion of the common iliac artery in performing cesarean section for patients with pernicious placenta previa and placenta accreta.Methods A total of five cases with ultrasound or MRI diagnosed pernicious placenta previa and placenta accreta were analyzed retrospectively.One of the cases was diagnosed Rh(-)blood type.Prophylactic temporary balloon implantation in bilateral common iliac arteries were carried out before cesarean section.Digital subtraction angiography ensured the position of balloon catheter and the catheter was fixed.The balloon was inflated immediately after the removal of the fetus.The balloon was removed at 6-8 hours after the cesarean section.The amount of blood loss,transfusion requirement,cesarean hysterectomy rate, and X-ray exposured time and dose during the procedure were recorded.Results Temporary balloon implantation in bilateral common iliac arteries in all five patients were obtained successfully.The blood loss was seen <500 mL in one patient and 500-1 000 mL in other four patients.Because of placenta implantation over depth of serosa and placenta percreta in one case,massive intractable hemorrhage occurred in short time,partial hysterectomy had to be carried out.The uterus was retained in other four cases.Conclusion The temporary balloon occlusion of the common iliac artery in performing cesarean section is a safe and effective technique,and it can reduce the amount of blood loss,transfusion requirement and secondary risk due to uncontrollable bleeding during surgery.

Objective To investigate the ultrasonic image characteristics of Zenker diverticulum(ZD).Methods The ultrasonographic features were summarized using the analysis confirmed by surgery pathology or X-ray imaging of the ZD in six eases.Results The diverticulum of Zenker located behind the thyroid gland in all the six cases,with five cases to the left and one to the right.The internal hyperechoic foci caused by air was found for all the six cases.The diverticulum wall appeared to be semicircular,narrow-band and acoustic halo low echo with an average thinckness of 0.53 mm.The Doppler sonogram of the six diverticulums showed no vascular signal.Conclusions The ZD had several unique characterisitics for identification by ultrasonic diagnosis,which included hyperechoic foci caused by air,internal changes of echo after drinking water and the special layered structure of the five-layer ZD wall.

Objective To synthesize 68 Ga-1,4,7,10-tetraazacyclododecane-N,N',N,N()-tetraacetic acid-D-Phe1-Tyr3-Thr8-octreotide (68Ga-DOTATATE) manually and automatically,validate its qualities in vitro,and evaluate its biodistribution in ICR mice and the microPET imaging in nude mice bearing pancreatic AR42J tumor.Methods 68Ga-DOTATATE was synthesized by automatic method using commercial metal isotope multifunctional module with strong cation exchange (SCX) column and by manual method.Both the products were measured for quality control.For the biodistribution study 5 groups of ICR mice were injected with 68Ga-DOTATATE(1.11 MBq) and executed at 10,30,60,120 and 240 min postinjection,respectively.The organs were weighted,and ％ ID/g was calculated.Nude mice bearing pancreatic AR42J tumor were intravenously injected with 3.7 MBq 68Ga-DOTATATE,and then microPET imaging was acquired at 10,30,60,120,18 and 240 min.Results 68Ga-DOTATATE could be successfully synthesized by the automatic and manual methods.Both the product injections were colorless and clear.The pH value was 6.5±0.1.For the products obtained from the two methods,the radiochemical purities were over 99％,and the products were stable for 4 h at room temperature.For the automatic method,68Ga-DOTATATE was synthesized within 30 min and with the radiochemical yield of (51.8±3.2)％ (time decay corrected).For the manual method,the time used for the synthesis was 20 min,and the labeling yield was over 99％.Three batches of the products were aseptic and pyrogen-free.In ICR mice,68Ga-DOTATATE was excreted by the kidney,and showed relatively high accumulation in the liver,spleen,pancreas and adrenal glands,while lower in the bone and soft tissue.The clearance from blood was fast with (4.41±0.81) ％ID/g at 10 min postinjection and (0.78 ± 0.32) ％ ID/g at 1 h.MicroPET imaging showed increased uptake of 68GaDOTATATE in the tumor tissues,and T/NT were 2.01±0.29(10 min),6.74±2.90(30 min) and 4.46±2.05 (60 min),respectively.Conclusions 68 Ga-DOTATATE could be successfully synthesized manually and automatically.The products reach to the specification of radioactive drugs and could be used as an attractive positron emitting radiotracer for detection of the somatostatin receptor-positive tumors.

Objective To detect the expression of aquaporin 1 in pancreas of rats with acute necrotizing pancreatitis (ANP) and to study the effect of Dachengqi decoction on it.MethodsOne hundred and sixty male SD rats were randomly divided into control group ( C group,n =32 ),ANP group ( n =32),Dexamethasone group (De group,n =32),Acetazolamide group (A group,n =32) and Dachengqi decoction group (DD group,n =32).ANP model was induced by retrograde injection of 5％ sodium taurocholate into the biliary and pancreatic duct.Rats in De group received dexamethasone (4 mg/kg) intravenously after ANP induction; while rats in A group received 1 ml acetazolamide via gastric lavage 2 h before ANP induction; rats in DD group received 2 ml Dachengqi decoction via gastric lavage 48,24,2h before ANP induction; rats in C group received laparotomy.Eight rats in each group were sacrificed at 3 h,6 h,12 h and 18h after induction of ANP models.Quantity of ascites and levels of serum amylases were measured.Pathological changes in pancreas tissue were detected by HE and electron microscope.Capillary permeability in pancreas tissue was detected by Evans Blue (EB) extravasations method.AQP1 expression in pancreas tissue was detected by real-time PCR and Western blotting.ResultsLevels of serum amylase in ANP group was significantly higher,and the pancreatic injuries were obvious ; the levels of serum amylase in De group and DD group was lower than that in ANP group,and the pancreatic injuries were attenuated.The levels of serum amylase in A group were higher than that in ANP group,and the pancreatic.injuries were more severe than that in ANP group.Six hours after ANP induction,the levels of EB in pancreas were (13.44 ±2.56),(126.35 ± 14.80),(86.31 ± 14.46),( 108.99 ± 15.07 ),(78.29 ± 16.85 ) mg/L In C group,ANP group,De group,A group and DD group,and the expression of AQP1 mRNA in pancreatic tissue was ( 170.07 ± 22.48 ) ％,( 83.93 ± 8.98 ) ％,( 117.09 ±10.70 ) ％,( 69.00 ± 8.98 ) ％,( 112.82 ± 11.79 ) ％ ; and the expression of AQP1 protein was 0.23 ± 0.06,0.10 ±0.02,0.32 ±0.03,0.13 ±0.02,0.45 ±0.04.The content of EB in ANP group was higher than that in C group,while the expression of AQP1 mRNA and protein in ANP group was significantly lower than that in C group (P ＜ 0.05 ).The content of EB in De group and DD group was significantly lower than that in ANP group,while the expression of AQP1 mRNA and protein was significantly higher than that in ANP group (P ＜ 0.05).ConclusionsAQP1 plays an important role in the pathogenesis of capillary endothelial barrier dysfunction in rats with ANP.Dachengqi Decoction can attenuate pancreatic injuries of rats by regulating the expression of AQP1.

Objective To explore the role of intracellular reactive oxygen species (ROS) in the senescence of bone marrow mesenchymal stem cells(BMSCs)in patients with systemic lupus erythematosus(SLE) and the underlying mechanisms that controls the intracellular ROS levels in vitro. Methods Human bone marrow aspirates were collected from iliac of eight donors and eight SLE patients and cultured in vitro.Morphological appearance of BMSCs at different passages was examined by inverted microscope. Nuclear size was measured by fluorescence microscope. BMSCs were monitored using the senescence associated β-galacto-sidase (SAβ-gal) assay to characterize senescence in vitro. The quantification of intracellular ROS production was detected by flow cytometry. Real-time PCR technique was used to determine the gene expressions of PI3K, KRas, NRas and FoxO3 at transcription level. The expression of FoxO3, phospho-FoxO3 (p-FoxO3),AKT and phospho-AKT (p-AKT) protein were determined by Western blot analysis. Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results There were no differences in morphology and nuclear size[(31.4±4.5) vs (28.2±4.8) μm, P=0.628] of BMSCs between SLE patients and normal controls.The percentage of SA β-gal positive BMSCs from SLE patients was higher than that from healthy controls [(31.8±9.0)% vs (12.4±0.7)%, P＜0.05]. Intracellular ROS levels of BMSCs from SLE patients increased more significantly than healthy donors in vitro (34600±9600 vs 17 958±5400, P＜0.05). No significant differences in the expression of PI3K, NRas, KRas and FoxO3 from SLE subjects were observed at mRNA levels compared with normal controls, though all showed a similar upward trend. The expression of p-FoxO3 and p-AKT of BMSCs from SLE patients increased significantly compared with healthy controls at protein levels.Conclusion These data suggest that BMSCs from SLE patients aged more quickly, with high SA β-gal activity and up-regulation of intracellular ROS, which is associated with up-regulation of p-FoxO3 and pAKT at protein levels. These results indicate that bone marrow mesenchymal cell senescence may be associated with the pathogcnesis of SLE by maintaining the lifespan of BMSCs.

ObjectiveTo investigate the effect of IκB kinase (IKK-β) on migration and proliferation of bone marrow mesenchymal stem cells(BMSCs) from patients with systemic lupus erythematosus (SLE).MethodsHuman bone marrow aspirates were collected from iliac of six donors and six SLE patients and cultured in vitro.Migration of BMSCs were observed by wound healing and transwell migration assays.Proliferation of BMSCs was quantified by cell counting kit-8 assay.Total RNA was extracted and reverse transcribed into complementary DNA.Real-time PCR technique was used to determine the gene expression of IKK-β at transcription level.The expression of IKK-β and phospho-IKK-β(p-IKK-β) protein were determined by Western blotting analysis.Statistical analysis was conducted with or Mann-Whitney rank test.Results① The migration rate of BMSCs from SLE patients(5.2±3.8)‰ were significantly reduced as compared with normal controls (7.0±2.9)‰(P＜0.05 ).The proliferation of BMSCs of SLE patients (0.21±0.49)was lower than that from healthy controls ( 1.00±0.35 )(P＜0.05 ).②) No difference in IKK-β3 mRNA expression between SLE ( 1.9± 1.4) subjects and normal controls (1.9±2.4) (P＞0.05).IKK-β protein expression of BMSCs from SLE patients (1.41 ±0.19) increased significantly compared with healthy controls (0.93±1.24) (P＜0.05).③ Inhibitor of IKK-β caused a significant increase in cell migration (3.3±1.6)‰ and proliferation (1.13±0.26) of BMSCs from SLE patients compared with untreated cells (2.3±1.1)‰ and (0.81±0.17),respectively (P＜0.05).ConclusionMigration and proliferation of BMSCs are significantly decreased in SLE patients.IKK-β may be involved in migration and proliferation of BMSCs.