Objective To compare the expressions of Beclin 1 and its 11 interacting proteins in subcutaneous fat and visceral fat,and to observe the expressions of these genes in 3T3-L1 adipocytes differentiation process.Methods In 3T3-L1 adipocytes,cells were harvested at day 0,2,4,6,8 in differentiation process,and then proteins or mRNAs were obtained and followed with Western blot and realtime PCR.Mice tissue proteins or mRNAs were detected with Western blot and realtime PCR.Results In 3T3-L1 adipocytes differentiation process,expressions of LC3,Beclin 1,and TNFAIP3 continued to rise,expression of BIRC5 was high to low,FEZ1 continued at a low level.In mice,expressions of LC3,Beclin 1,YWHAQ,FEZI,BAD,WAC,TNFAIP3,and GOPC in visceral fat were significantly higher than those in subcutaneous fat,while expression of SLAMF1was higher in subcutaneous fat.Conclusion Autophagy plays an important role in 3T3-L1 differentiation process and may be tissue-specific in visceral fat and subcutaneous fat.

Tumor microenvironment (TME) plays a key role in the development and progression of tumors, such as pro?moting local drug resistance, immune escape, and distal metastasis. According to the TME of different individuals, accurate evaluation and selection of clinical medication can effectively control the malignant transformation of carcinoma in situ and metastatic cancer. At present, the main method to treat cancer is chemotherapy, TME can regulate the reaction of the tumor cells to the standard chemotherapy and target drug therapy, so the combination of the targeted TME therapy and chemothera?py will achieve better clinical efficacy. In this review, we summarized the mechanisms of TME in breast cancer, including ex?tracellular matrix, carcinoma-associated fibroblasts, carcinoma-associated macrophages, regulatory T cells and bone marrow mesenchymal stem cells, which providing a theoretical basis for the development of TME targeted therapy.

Objective To explore the expressions of Smad4 and estrogen receptor (ER) and their interrelation,and the relationship with the clinicopathological features of breast cancer.Methods The immunohistochemical SP method was used to detect the expressions of Smad4 and ER in 50 case of invasive cancer,12 cases of carcinoma in situ and 15 cases of normal breast tissues.The differences in different clinical stages,differentiation degrees and nodal metastases were analyzed.The correlation between Smad4 and ER was explored.Results The positive expression rate of Smad4 in invasive cancer was 52.00％,which lower than that in normal breast tissue (93.33％),with a significant difference (x2 =8.329,P =0.004),positive expression rates of ER were 60.00％ and 40.00％ respectively,with no significant difference (x2 =1.868,P =0.172).The positive expression rates of Smad4 in carcinoma in situ and invasive cancer were 75.00％ and 52.00％ respectively,with no significant difference (x2 =2.082,P =0.149).The positive expression rates of ER were 58.33％ and 60.00％ respectively,with no significant difference (x2 =0.011,P =0.916).The positive expression of Smad4 was related to the TNM stage (x2 =6.392,P =0.011) and the lymph node metastasis (x2 =6.738,P =0.009),but it was not associated with the histologic grade (x2 =0.542,P =0.462).The positive expression of ER was related to the lymph node metastasis (x2 =4.133,P =0.042) and histologic grade (x2 =5.357,P =0.021),but it was not associated with the TNM stage (x2 =1.159,P =0.282).There was positive correlation between Smad4 and ER in breast cancer tissue (r =0.263,P =0.032).Conclusion Smad4 is expressed at lower level in breast cancer than in normal breast tissue.The expressions of Smad4 and ER are related to the different clinicopathological features of breast cancer with positive correlation.

Objective To evaluate the application value with Energy Spectrum CT multi -parameter quanti-tative value in differentiating pulmonary occupying lesions (>0.5cm in diameter).Methods Those were retrospec-tively analyzed on 58 cases in pulmonary occupying lesions performed spectrum dual -phase enhanced CT scan and confirmed by pathology,including malignant group 48 cases;10 cases of benign group.Iodine content(IC)was meas-ured in arterial phase(AP)and venous phase (VP)in iodine based on the image and the level of the aorta,and nor-malized iodine concentrations were calculated in the two phases lesions compared with aorta,and the difference between the two normalized iodine concentrations ICD (ICD =NICvp -NICap).To measure the CT value of lesions on 50keV and 100keV energy image,and calculate the energy attenuation curve slope,namely |Hu (100 kev -50kev)/Hu50 |.The differences of NIC,curve slope,and ICD were compared between benign and malignant pulmonary occupying lesions using independent sample t test method.Results In two phase scan,NIC,curve slope of the venous phase and ICD in malignant group were significantly higher than the benign group,(NIC,curve slope of the venous phase and ICD of the malignant group:NICap:0.180 ±0.051,NICvp:0.463 ±0.086,1.696 ±0.475,ICD:0.284 ± 0.071;NIC,curve slope of the venous phase and ICD of the benign group:NICap:0.123 ±0.062,NICvp:0.290 ± 0.119,1.169 ±0.582,ICD:0.166 ±0.073,),but there was no significant difference between the patients with benign and malignant lesions in curve slope of the arterial phase.Conclusion Energy spectrum CT dual -phase enhanced scan can differentiate the nature of benign and malignant pulmonary occupying lesions,and has certain clinical application value.

Objective To establish a comprehensive,simple,and effective scoring model for predicting the recurrence risk after endovascular embolization of intracranial aneurysms in order to assess the possibility of recurrence and to provide guidance for the selection of surgical protocols and postoperative management.Methods From May 2012 to May 2014,434 patients (441 aneurysms) with intracranial aneurysm treated with endovascular embolization at the Department of Neurosurgery,Changhai Hospital,the Second Military Medical University were enrolled retrospectively,and they were used as a modeling group.After modeling,109 patients (109 aneurysms) were used as a validation group.In the modeling cohort,a predictive scoring model of recurrence risk was established according to the results of multivariate logistic regression analysis;the model was validated in the validation cohort.According to the scoring model of the modeling group,the scoring table of best cut-off value of the receiver operating characteristic (ROC) curves was divided into a low-risk and a high-risk of recurrence.The recurrence risk score model was compared with the North America aneurysm recanalization stratification scale (ARSS) model,and Raymond grade.Results Multivariate logistic regression analysis showed that the 3 factors included in the scores and finally,a established scoring model of recurrence risk prediction were non-stent assisted embolization (1 point),Raymond grade ≥Ⅱ (1 point),and the size of aneurysm (aneurysm >25 mm[3 points)],aneurysm 10-25 mm[1 point],and aneurysm <10 mm[0 point]).The validation indicated that the scoring system had higher predictive value (AUC=0.738,95%CI 0.641-0.834,P<0.05) and goodness of fit (Hosmer-Lemeshow χ2=2.109,P=0.146).The scoring table was further divided into the low-risk recurrence (0-1 point) and high-risk recurrence (2-5 points),its sensitivity was 72.73% (48/66) and specificity was 68.80% (258/375).The predictive ability of the aneurysm recurrence risk score model was similar to that of the ARSS score (χ2=0.54,P=0.462),and it was better than the Raymond grade (χ2=15.10,P<0.01).Conclusion The established simple aneurysm recurrence risk predicting score model in this study may accurately predict the recurrence of aneurysms,however,a multicenter,large sample prospective study is needed for further validation.

The occurrence and progression of cervical cancer are influenced by gut flora. Patients with cervical cancer have different gut flora from healthy women, and the detection and evaluation of gut flora can help in the diagnosis, immunotherapy, assessment of radiotherapy efficacy and prognosis prediction of cervical cancer. Regulation of gut flora is of great clinical value in enhancing response to anti-tumor therapy, improving patients' quality of life and improving prognosis.

Immune checkpoint inhibitors in the treatment of head and neck squamous cell carcinoma (HNSCC) has shown significant clinical benefit. Some studies have shown that radiotherapy combined with immunotherapy can produce synergistic effects, and several phase Ⅰ/Ⅱ clinical trials have suggested that radiotherapy combined with immunotherapy has good safety and preliminary efficacy benefits in locally advanced HNSCC. However, the timing of combination therapy, the selection of radiotherapy dose/fractionation mode and patients are still unclear. This article further discusses the synergistic mechanism, clinical research status and challenges of radiotherapy combined with immunotherapy in the treatment of HNSCC, aiming to guide the clinical practice and improve the prognosis of HNSCC patients.

Objective To investigate the clinical efficacy of magnetic compression anastomosis for congenital esophageal atresia and stenosis.Methods The retrospective and descriptive study was conducted.The clinical data of 4 children who underwent magnetic compression anastomosis for congenital esophageal atresia and stenosis in the Northwest Women and Children's Hospital from December 2017 and February 2019 were collected.There were 2 males and 2 females.The children were aged 11 days,7 days,5 days,and 3 years,respectively.The children underwent magnetic compression anastomosis.Observation indicators:(1) surgical and postoperative situations;(2) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect food intake and complications of children up to May 2019.Measurement data with normal distribution were represented as Mean±SD,and measurement data with skewed distribution were represented as M (range).Results (1) Surgical and postoperative situations:four children underwent magnetic compression anastomosis successfully.Of the 4 children,3 with esophageal atresia underwent open tracheoesophageal fistula repair and endoscopeassisted magnetic compression anastomosis,and 1 with congenital esophageal stenosis underwent endoscopic gastrostomy combined with magnetic compression anastomosis.The operation time of 4 children was (2.3±0.9) hours.The length of esophageal blind ending in the 3 children with esophageal atresia and length of esophageal stenosis were in the children with esophageal stenosis 30-35 mm and 8 mm.Four children has good magnet apposition,and time of postoperative magnet removal was (29± 10)days.Three children with esophageal atresia had oral removal of magnet,and 1 with esophageal stenosis had magnet removed by gastrostomy.One child complicated with postoperative fistula and anastomotic stenosis was cured by unobstructed drainage and nutritional support treatment.The duration of postoperative hospital stay was (39± 10)days.(2) Follow-up:4 patients were followed up for 3-17 months,with a median time of 10 months,and restored to oral intake after oral removal of magnet and removal of magnet by gastrostomy on the days 14-36 postoperatively.One child was detected anastomotic stenosis by esophagography at the postoperative 3 months,and was improved after esophageal dilatation.The other 3 children recovered to normal connectivity of esophagus postoperatively and maintain unobstructed.Four children had normal eating,without dysphagia or other serious complications.Conclusion Magnetic compression anastomosis is safe and feasible for congenital esophageal atresia and stenosis,with good short-term efficacy.

Objective:To investigate the clinical, pathological and genetic features of 3 cases of limb-girdle muscular dystrophy 2A (LGMD2A) caused by non-canonical splice site mutations in the CAPN3 gene. Methods:For the 3 LGMD2A patients admitted to Qilu Hospital of Shandong University from July 2016 to July 2018 were selected as the subjects. Clinical data were collected, whole exome sequencing was conducted, and the candidate variants were verified by Sanger sequencing. Total RNA was extracted from the skeletal muscle tissue of 3 probands and effects of splicing mutations on pre-mRNA splicing in the CAPN3 gene were verified by reverse-transcription polymerase chain reaction. Total protein was extracted from the muscle tissue of the probands and expression level of calpain 3 protein was detected by Western blotting. Results:All the 3 probands presented muscle weakness in upper and lower limbs, and muscle weakness in proximal limbs was more severe. Muscle biopsies all indicated myogenic impairment. Genetic sequencing showed proband 1 carried compound heterozygous c.2185-14T>G and c.2305C>T (p.R769W) mutations in the CAPN3 gene, proband 2 carried compound heterozygous c.1193+30G>A and c.2069_2070delAC (p.H690Rfs *9) mutations, and proband 3 carried homozygous c.1194-9A>G mutations in the CAPN3 gene. Splicing assay showed the c.2185-14T>G mutation located in intron 20 induced retention of the entire intron 20, the c.1193+30G>A mutation in intron 9 induced retention of the first 31 nucleotides of intron 9, and the c.1194-9A>G mutation in intron 9 induced retention of the last eight nucleotides of intron 9. Western blotting revealed deficiency of calpain 3 protein in skeletal muscle of proband 1 and proband 2. Conclusions:The clinical manifestation of LGMD2A is muscle weakness predominantly in proximal limbs, and the muscle pathology is mostly characterized by myogenic impairment.Moreover, aberrant splicing of pre-mRNA caused by non-canonical splice site mutations plays a pathogenic role in this disease.

Objective:To verify the ability of the newly developed ultra-high resolution CT (U-HRCT) for displaying the fine bony anatomy of temporal bone.Methods:The cone-beam CT architecture was used on U-HRCT. The focus size of X-ray generator was 0.27 mm × 0.29 mm, the rated tube voltage was 60-100 kV, and the unit size of flat panel detector was 0.074 8 mm×0.074 8 mm. From October to December 2019, 16 adult head specimens (32 temporal bones, provided by Huanghe University of Science and Technology) were imaged using U-HRCT. The scanning mode was small-field-of-view and high-resolution scanning for unilateral temporal bone. The scanning parameters were: voltage 100 kV, current 3.5 mA, rotation time 40 s, reconstructed field of view 65 mm×65 mm, voxel size 0.1 mm×0.1 mm×0.1 mm, layer number 370, slice thickness 0.1 mm, and slice interval 0.1 mm. The ultimate spatial resolution of the system was detected using a phantom with line pair card, after the scanning with the same parameters. Through multi-planar reconstruction and minimum intensity projection method, a total of 6 anatomical positions of 4 structures (stapes footplate, cochlear axis bottom, vestibular aqueduct internal orifice and isthmus, cochlear aqueduct internal orifice and auditory sac segment) were scored with 1-3 points. The Wilcoxon test was used to compare the difference of bilateral scores of each structure.Results:The phantom test results showed that the ultimate spatial resolution of the system was ≥4.0 lp/mm. The scores of stapes footplate, cochlear axis bottom and vestibular aqueduct internal orifice were ≥2 points, with a display rate of 100%. The display rates of vestibular aqueduct isthmus, cochlear aqueduct internal orifice and auditory sac segment were 87.5% (28/32), 71.9% (23/32) and 53.1% (17/32), respectively. There were no significant differences in left and right scores of all anatomical structures ( P>0.05). Conclusion:The newly developed U-HRCT has good display ability for fine bony anatomy of temporal bone and has great clinical application potential.