Objective:To investigate the ultrasonographic characteristics and risk factors of breast cancer internal mammary lymph node (IMLN) metastasis.Methods:A retrospective analysis of 296 first diagnosed breast cancer patients in the Fourth Hospital of Hebei Medical University from March 2010 to May 2020. IMLN was divided into metastatic group (236 cases) and non-metastatic group (60 cases) based on pathology. Chi-square test and independent sample t test were used to analyze the ultrasound characteristics of IMLN metastasis and factors related to metastasis. ROC curve analysis of IMLNs were plotted to obtain the diagnostic thresholds and their sensitivity and specificity.Univariate and multivariate Logistic analysis was used to analyze the risk factors of IMLN metastasis. Results:①The appearances of IMLN in ultrasound were normal type, thickened-cortex type, unclear hilus structure type and thickened-nodular soft tissue type. ②In the two groups, the differences in IMLN long diameter, thickness diameter, number, and lymphatic hilum structure type were statistically significant (all P<0.05), and there were no significant differences in IMLN long diameter/thickness diameter and IMLN blood supply (all P>0.05). ③The long diameter threshold of IMLN for diagnosis of metastasis was 10.5 mm, the are under the ROC curve(AUC) was 0.825, with sensitivity of 58.5% and specificity 93.3%; thickness and diameter threshold was 4.5 mm, AUC was 0.790, with sensitivity 66.9% and specificity 75.0%. The sensitivity and specificity of the diagnosis of long-diameter combined structure type were 56.3% and 93.3%, respectively; the sensitivity and specificity of the diagnosis of thick-diameter combined structure type were 64.8% and 81.7%, respectively. The cortical thickness threshold was 1.9 mm, and the diagnostic sensitivity and specificity were 91.9% and 86.7%, respectively. ④The risk factors of IMLN metastasis inculded univariate analysis showed tumor length, tumor volume, axillary lymph node long diameter, axillary lymph node metastasis, and clavicle lymph node metastasis. There was a statistically significant difference in the pathology of the lower lymph nodes between the two groups ( P<0.05). Multivariate analysis showed that the long diameter of the tumor and the metastasis of the axillary lymph nodes were independent risk factors of IMLN metastasis. Conclusions:The metastatic IMLN mostly manifest as no lymphatic hilum structure or cortical thickening (≥1.9 mm), and multiple IMLN can help diagnose metastasis.Ultrasound can better assess breast cancer IMLN metastasis, and the diagnostic efficiency of IMLN long-diameter combines structure type is higher.Independent risk factors for IMLN metastasis include tumor size and axillary lymph node metastasis.

ObjectiveToinvestigate the prevalence, awareness and risk factors of chronic kidney disease (CKD) among community adult population in Shanghai, China, in order to provide early diagnosis and treatment of CKD, and informations for national health policy makers.MethodsTwo thousand five hundred and ninety six residents (≥ 18 years old) were randomly selected from community population in Changning district of Shanghai, China. They were interviewed and tested for albuminuria -morning spot urine albumin to creatinine ratio [ACR, abnormal: ≥ 17 mg/g (male), ≥25 mg/g (female)], reduced renal function-estimated GFR by abbreviated MDRD equation [abnormal: <60 ml ·rain-1 (1.73 m2)-1] and hematuria-morning spot urine dipstick confirmed by urine microscopy. The associations among demographic characteristics, healthy characteristics (e.g. diabetes and hypertension) and indicators of kidney damage were examined. The investigators and neighborhood committee were well trained. Those who had semiquantitative positive were detected again by albuminuria-morniag spot urine albumin to creatinine ratio after three months. ResultsTwo thousand five hundred and fifty four residents with complete data were enrolled in the study. Albuminuria was detected in 6.3% of subjects, reduced renal function in 5.8%, hematuria in 1.2%. Approximately 11.8% of these subjects had at least one indicator of kidney damage. The awareness rate of CKD was 8.2%. The Logistic regression model showed that hyperuricemia, nephrolithiasis, anemia, diabetes, central obesity, hypertension and age contributed to the development of CKD. ConclusionsThe prevalence of CKD in community adult population in Shanghai is 11.8%, And the awareness rate of CKD is 8.2%. Hyperuricemia, nephrolithiasis, anemia, diabetes, central obesity, hypertension and age are risk factors of CKD.

Objective:To determine the influences of structural changes after valgus impacted femoral neck fracture on hip range of motion (ROM) so as to provide evidence for clinical judgment of whether reduction is necessary or not in the internal fixation of such fractures.Methods:1. 3D reconstructions of the CT hip scans were performed for the 73 patients who had been treated at Department of Orthopaedic Surgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University for valgus impacted femoral neck fractures from January 2019 to April 2019.The femoral neck-shaft angle, anteversion angle, femoral offset, axial alpha angle, lateral center edge angle (LCEA), anterior center edge angle (ACEA) and center displacement were measured and compared between the affected and healthy sides to determine the influences of the fracture on the above indexes. 2. Hip motions (flexion and MIR-90°) were simulated on bilateral sides to determine the influences of structural changes after fracture on hip ROM using stepwise regression and Logistic regression. 3. The distribution of femoral-acetabular contact points on the femoral side was observed in simulation of hip flexion to detect the potential area for femoracetabular impingement (FAI) induced by the fracture displacement.Results:1. The valgus impacted femoral neck fractures had significant influences on femoral neck-shaft angle, anteversion angle, femoral offset and axial alpha angle. Compared with the healthy side, on average, the femoral neck-shaft angle increased by 5.1°, anteversion angle decreased by 6.5°, femoral offset decreased by 8.2 mm and axial alpha angle increased by 9.7° on the affected side, showing significant differences ( P<0.05).The displacements of the femoral head center averaged 9.2 mm. There was no significant difference in LCEA or ACEA between the affected and healthy sides ( P>0.05). 2. Compared with the healthy side, on average, the simulated hip flexion decreased significantly by 27.0° and the hip MIR-90° decreased significantly by 20.3° on the affected side after fracture ( P<0.05). Regression analysis showed that femoral anteversion angle, ACEA and displacement of the femoral head center had a significant influence on hip ROM, especially the anteversion angle. When the anteversion angle decreased by more than 7.1°, the hip flexion would decrease by at least 20%. 3. The points of FAI distributed more widely on the fracture side. Compared with the healthy side, the impact points extended outward and upward in hip flexion and extended inwardly in hip MIR-90° on the affected side. Conclusions:After a valgus impacted femoral neck fracture, if the femoral anteversion angle has been decreased by more than 7.1°, the hip ROM can be greatly influenced and the points of FAI can be distributed more widely. Therefore, reduction should be recommended before internal fixation of the fracture.

Purpose To explore the expression of autoph-agy-related 7(ATG7)in breast cancer and its effect on the breast cancer development.Methods Immunohistochemistry(IHC)was used to detect ATG7 protein expression in breast cancer tissues and the relationship between ATG7 and clinico-pathological features was analyzed.ShRNA was used to interfere with the expression of ATG7 in breast cancer cell line MCF-7.Puromycin was used to screen for stably transfected cells and Western blot was used to detect transfection efficiency.The effect of ATG7 knockdown cells on proliferation ability was de-tected by CCK8 and clone formation experiments.The effect of ATG7 knockdown cells on tumorigenicity in vivo was detected by subcutaneous tumor formation experiment in nude mice.Results IHC showed that ATG7 expression in breast cancer tissues was mainly localized in cytoplasm,and its expression was significant-ly correlated with tumor size and Ki67 expression(P＜0.05).ATG7-shRNA significantly interfered with ATG7 expression in breast cancer cells MCF-7.CCK8 and clone formation experi-ments showed that ATG7 knockdown promoted the cell prolifera-tion compared with the control group.The experiment of subcu-taneous tumor formation in nude mice showed that the tumor for-mation ability of mice was significantly increased after ATG7 knockdown compared with the control group.Conclusion ATG7 may inhibit the proliferation capacity of breast cancer and could be a potential target for breast cancer therapy.

Objective: To establish fluorescence quantitative polymerase chain reaction (PCR) to detect integrated HIV DNA in peripheral blood mononuclear cells. Methods: A total of 30 HIV-seropositve individuals were enrolled in this study, including 10 subjects with a detection limit of 20 copies/ml of plasma, 10 patients with drug resistance and 10 patients with no history of antiretroviral therapy (ART). Cultivated ACH2 cells carried a single copy of the integrated HIV genome. We have built pMD19T-CD3 plasmid and calculated the copy number. We used oligonucleotides ULF1 specific for the long terminal repeats (LTR) regions and two oliligonucleotides specific for human Alu sequences to pre-amplified the integrated HIV DNA. Samples and serial dilutions of ACH2 cells were all pre-amplified, the products of which were used for the second round fluorescence amplifications. The Lambda T primers, UR2 primers and HIV Taqman probes were used for second round amplifications in integrated HIV DNA assay. The CD3IN5 primers, CD3IN3 primers and CD3 Taqman probes were used for CD3 quantification. Results: Serial 5-fold dilutions of the plasmid were used as standards for CD3 gene quantifications. The equation of the linear regression was y=-2.731x+ 43.01(R²=0.953). The cellular input was quantified by number of human genome equivalents (CD3 gene, 2 copies/cell). The copies of ACH2 cells was 4.271×10^4, which was the cellular input of the ACH2 cells. Serial 5-fold dilutions of ACH2 cells were used to generate a standard curve for the integrated HIV DNA assays. The equation of the linear regression was y=-3.146x+ 39.11 (R²=0.968). The ratio between the number of copies of the integrated HIV DNA form and the number of cells (CD3 copies) was calculated to obtain the frequency of cells. Based on the test between different groups, each group had no difference (P> 0.05). Conclusions This method presented advantage in detection of the lower copies of virus. It hinted that the current antiretroviral therapy can not effectively attack against latent viral reservoirs. It also provided a basis for new therapeutic intervention.

Objective To explore the effect of glioma stem cells with high expression of protein tyrosin phosphatase receptor type Z1 (PTPRZ1 )on the phenotypic polarization and phagocytosis of tumor-associated macrophages and its regulatory mechanism.Methods GSCs and non-stem tumor cells (NSTCs) were screened out from human glioblastoma (GBM) specimens using flow cytometry,and the PTPRZ1 expression in paired GSCs and NSTCs were detected.Human peripheral blood mononuclear cells (PBMC)-derived CD14+monocytes were exposed to the conditioned medium from glioma cells or recombinant chemokine C-C motif ligand 20 (CCL20)for TAM polarization.Stable PTPRZ1 knockout GSCs (PTPRZ1-KO GSCs) were constructed using CRISPR/Cas9. TAM phagocytosis to GSCs,NSTCs,PTPRZ1-Control GSCs (PTPRZ1-Ctrl GSCs)and PTPRZ1-KO GSCs and the expression of immunosuppressive phenotype (M2) polarization marker CD163 were examined using flow cytometry.Differentially expressed genes (DEGs ) between paired GSCs and NSTCs were determined using a bulk RNA-sequencing dataset (GSE54791 )from Gene Expression Omnibus (GEO).A gene set informing worse outcome of patients with GBM was generated using The Cancer Genome Atlas (TCGA)-GBM cohort.By intersecting the aforementioned gene set with the gene set that encodes for human membrance proteins,the PTPRZ1 gene is obtained.Gene set enrichment analysis (GSEA)was used for pathway enrichment analysis to compare the differentially regulated pathways between GBMs with high or low PTPRZ1 expression.Bulk RNA sequencing,qRT-PCR and Western blotting were used to identify the DEGs between PTPRZ1-KO GSCs and PTPRZ1-Ctrl GSCs.Results GSCs were more capable of escaping from TAM phagocytosis than NSTCs (P<0.05 )and had specifically up-regulated PTPRZ1 expression.PTPRZ1-KO significantly suppressed GSCs escaping from TAM phagocytosis (P<0.01 ). GBMs with high PTPRZ1 expression showed significant inhibition of pathways mediating phagocytosis (P<0.05).The expression of CCL20 as a M2 TAM polarization chemokine was significantly down-regulated in PTPRZ1-KO GSCs (P<0.05 ).Treatment with recombinant CCL20 up-regulated the expression of CD163 as a M2 TAM marker in TAM.Conclusion PTPRZ1+GSCs mediate M2 TAM polarization and inhibit TAM phagocytosis,which may be related to the up-regulation of CCL20 in PTPRZ1+GSCs.

BACKGROUNDBone marrow hematopoietic function suppression is one of the most common side effects of chemotherapy. After chemotherapy, the bone marrow structure gets destroyed and the cells died, which might cause the hematopoietic function suppression. Heme oxygenase-1 (HO-1) is a key enzyme of antioxidative metabolism that associates with cell proliferation and resistance to apoptosis. The aim of this study was to restore or resist the bone marrow from the damage of chemotherapy by the HO-1 expression of mouse mesenchymal stem cells (mMSCs) homing to the mice which had the chemotherapy-induced bone marrow suppression.

METHODSOne hundred and sixty female Balb/c mice (6-8-weeks old) were randomly divided into four groups. Each group was performed in 40 mice. The control group was intraperitoneally injected for 5 days and tail intravenously injected on the 6th day with normal saline. The chemotherapy-induced bone marrow suppression was established by intraperitoneally injecting cyclophosphamide (CTX) into the mice which performed as the chemotherapy group. The mMSCs were tail intravenously injected into 40 chemotherapically damaged mice which served as the mMSCs group. The difference between the HO-1 group and the mMSCs group was the injected cells. The HO-1 group was tail intravenously injected into the mMSCs that highly expressed HO-1 which was stimulated by hemin. The expression of HO-1 was analyzed by Western blotting and RT-PCR. Cell proliferation was measured using the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Histopathologic examinations were performed 1 week after injection.

RESULTSCompared with the control group, the expression levels of HO-1 mRNA and protein were significantly higher in the HO-1 group (all P < 0.05), even obviously than the mMSCs group. CTX treatment induced apoptosis and inhibited proliferation. After injected, the white blood cell (WBC), red blood cell (RBC) and platelet (PLT) declined fast and down to the bottom at the 7th day. The bone marrow structure was destroyed incomplete. In vitro, the survival rate of cells in chemotherapy group was less than 50% after 24 hours. In contrast, mMSCs could do a favor to the cellular cleavage and proliferation. They slowed down the cell mortality and more than 50% cells survived after 24 hours. The effects of blocking apoptosis and bone marrow recovery could be more effective in the HO-1 group. In the HO-1 group, it had observed that the bone marrow structure became complete and the hemogram closed to normal at 7th day.

CONCLUSIONSHO-1 played an important role in promoting the recovery of CTX-induced hematopoietic damage. We suggest that HO-1 is able to restore the functions of chemotherapy-induced hematopoietic damage.

Animals ; Apoptosis ; drug effects ; Blood Platelets ; drug effects ; Blotting, Western ; Bone Marrow ; drug effects ; enzymology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclophosphamide ; toxicity ; Erythrocytes ; drug effects ; Female ; Heme Oxygenase-1 ; genetics ; metabolism ; Leukocytes ; drug effects ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; enzymology ; physiology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVE： To study the effects of Tiaopi huxin prescription （TPHXP） on the atherosclerosis （AS） of ApoE－/－ mice， and to investigate its mechanism. METHODS： Forty male ApoE－/－ mice were divided into blank group， model group， simvastatin group （positive control， 5 mg/kg） and TPHXP low-dose and high-dose groups （50， 150 mg/kg）， with 8 mice in each group. Except that blank group was given common diet， other groups were given high-lipid diet to induce AS model. After modeling， administration groups were given relevant medicine intragastrically， and blank group and model group were given constant volume of normal saline intragastrically， once a day， for consecutive 12 weeks. After last medication， the serum levels of TC， TG， LDL-C and HDL-C were determined by spectrophotometry. The serum level of NO was detected by nitrate reduction method. The serum levels of IL-6 and VCAM-1 were determined by ELISA. After separating thoracic aorta， HE staining was used to observe the formation of plaque in the thoracic aorta of mice in each group， and the corrected plaque area was calculated. Western blotting was conducted to determine the expression of NF-κB p65， Cav-1 and eNOS. RESULTS： Compared with blank group， the serum levels of TC， TG， LDL-C， IL-6 and VCAM-1 were increased significantly in model group， while the levels of HDL-C and NO were decreased significantly （P＜0.01）. The plaque of thoracic aorta was obvious and the corrected plaque area were increased significantly （P＜0.01）. The relative expression of NF-κB p65 and Cav-1 were increased significantly， while the relative expression of eNOS was decreased significantly （P＜0.01）. Compared with model group， the serum levels of TC， TG and LDL-C in administration groups， the serum levels of IL-6 and VCAM-1 in simvastatin group and TPHXP high-dose group were decreased significantly， while the serum levels of HDL-C and NO were increased significantly in administration groups （P＜0.05 or P＜0.01）. In administration groups， the plaques of thoracic aorta were reduced and the corrected plaque area was decreased significantly （P＜0.05 or P＜0.01）； the relative expression of NF-κB p65 and Cav-1 were decreased significantly， while the relative expression of eNOS was increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： TPHXP can regulate the level of blood lipid， decrease the level of inflammatory factors and inhibit the formation of AS plaque， the mechanism of which may be associated with inhibiting Cav-1/NF-κB pathway.