Objective To evaluate the damage caused by high intensity ultrasound(HIU)on tumor micro- vessels.Methods Rabbit models of the VX_2 tumor were set up.The target hepatic carcinomas were treated with HIU,and the results were observed using hematoxylin-eosin(HE)staining,vascular endothelial cell biotinylated-ulex europaeus agglutininiⅠ(UEAI)immunohistochemical staining,tumor micro-vessel counts,and electron microscopy. Results Histological examination through HE staining indicated that HIU induced carcinoma vascularities with whole tumor tissue coagulative necrosis.The UEAI immunohistochemical staining of the target lesions treated by HIU was ne- gative,and complete tumor micro-vessel uhrastructure damage was observed under the electron microscope.Conclu- sion HIU can damage tumor micro-vessels thoroughly,in such way that it may inhibit tumor growth and metastasis.

OBJECTIVETo compare differences between Crowe IV developmental dysplasia of the hip (DDH) with secondary acetabulum and Crowe IV DDH without secondary acetabulum,and determine whether it is necessary to divide Crowe IV DDH into two subtypes.

METHODSFrom June 2007 to May 2015,145 hips of 112 Crowe N patients who underwent total hip arthroplasty (THA) using S-ROM stem were divided into two groups: secondary acetabulum formaton group (group A) and no secondary acetabulum formaton group (group B). In group A,there were 12 females, 96 males,with an average age of (39.38 ± 11.19) years old. In group B, there were 2 females, 35 males, with an average age of (38.19 ± 10.92) years old. All the patients were evaluated by using Harris Hip Score. Radiographic evaluations were made preoperatively and during follow up. The differences between two groups were compared on dislocation height, canal flare index (CFI), subtrochanteric shortening osteotomy (SSTO) usage, pre- and post-operation Harris scores, complications.

RESULTSThe dislocation height for group A was (4.74 ± 1.57) cm, while the dislocation height for group B was (3.12 ± 1.15) cm. Significantly difference was detected between two groups. The CFI for group A was 2.69 ± 0.68, while the CFI for group B was 3.42 ± 0.79, and the significantly difference was detected between two groups. Harris scores were totally improved from 58.18 ± 15.67 preoperatively to 91.20 ± 3.79 post-operatively and the difference was significant. Pre-operative Harris scores was 58.1 ± 15.3 in group A, 58.3 ± 16.9 in group B. Post-operative Harris scores was 91.0 ± 4.1 in group A, 91.0 ± 5.1 in group B. No significant difference was found on Harris scores between A and B preoperatively and post-operatively. Complications of 4 cases peri-prosthesis fracture, 4 cases dislocation and 4 cases nerve injury occur in group A; While only one case dislocation and one case nerve injury occur in group B. No statistical significance was detected.

CONCLUSIONCrowe IV DDH with secondary acetabulum is significantly different from Crowe IV DDH without secondary acetabulum on dislocation height and femoral morphology, which causes the different selections of surgical techniques (SSTO usage or not). These important differences in fundamental parameters indicate the necessity to further divide Crowe IV DDH into IVA and IVB two subtypes.

Adolescent ; Adult ; Aged ; Female ; Hip Dislocation, Congenital ; classification ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; therapy

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells (BCSCs). BCSCs were enriched by using serum-free medium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and flow cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a promising therapeutical approach for breast cancer.

AIMTo investigate the water-soluble steroidal saponins in total saponin from Dioscorea nipponica Makino and look for new active compounds.

METHODSThe compounds were isolated with silica gel, PTLC and HPLC, and their structures were elucidated by acid hydrolysis, physical and chemical data and spectral analysis (IR, NMR, MS, HMQC, HMBC) as well as chemical correlations.

RESULTSThe two steroidal saponins (water-insoluble saponin and water-soluble saponin) were isolated from the total saponin of Dioscorea nipponica Makino. The structures were elucidated as diosgenin 3-O-[alpha-L-rhamnopy ranosyl (1-->2)-[beta-D-glucopyranosyl(1-->3)]]-beta-D-glucopyranoside (I), diosgenin 3-O-[alpha-L-rhamnopyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl (1-->4)]-beta-D-glucopyranoside (II).

CONCLUSIONCompound II is a new steroidal saponin and firstly isolated from Dioscorea nipponica Makino. It was named as dioscin Dc.

Dioscorea ; chemistry ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Saponins ; chemistry ; isolation & purification

OBJECTIVETo summarize experience of using screws and cement to rebuild tibial bone defect in primary total knee arthroplasty (TKA) and to discuss the relationship between the number of required screws and the severity of tibial bone defects.

METHODSFrom July 2009 to May 2015, 34 patients (40 knees) with varus knees underwent TKA, and the screw and cement technique was used to rebuild medial tibia plateau during operation. There were 8 males (8 knees) and 26 females (32 knees), and the average age was (65.00 +/- 7.25) years old (ranged,55 to 82 years old). One to 6 screws were used in each case. Extension stems were used in 2 cases (4 and 5 screws was used respectively). The area percentages of the bone defects measured as defect area/tibia plateau area, depth of each defect, the number of screws needed in each case, were all used to determine the relationship between the number of screws and the area percentage in certain depth of bone defect by statistic methods, as well as the relationship between screw number and defect depth.

RESULTSAll the patients were followed up and the average duration was 24 months (ranged, 1 to 72 months). The average preoperative HSS score was 43.33 +/- 6.11 (ranged, 32 to 51 scores). Whereas the average postoperative HSS score was 92.15 +/- 4.64 (ranged,83 to 96 scores). The preoperative individual scores including pain, function, activity, nuscle strength, flexion deformity and stability were all improved compared with preoperation,and the differences were statistically significant. All the patients received normal alignment postoperatively, femoraltibial angle was improved from (167.00 +/- 6.39) degrees preoperatively to (175.00 +/- 2.69) degrees postoperatively, the tibial angle was improved from (78.09 +/- 4.51) degrees preoperatively to (88.75 +/- 1.24) degrees postoperatively. Both area percentage and depth of bone defect in a fitting Ologistic model had a significant statistical relationship with the screw number, and a rectangular coordinate system could be formed according to the relationship.

CONCLUSIONScrews and cement technique is a simple, safe and convenient method to rebuild tibial bone defects in primary TKA and its short-term and midterm effect are both reliable. During opera- tion, according to the rectangular coordinate system, the screw number needed in the operation can be inferred form th area and depth of tibia defect, which could have a guiding function in surgery.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; instrumentation ; methods ; Bone Screws ; utilization ; Female ; Fracture Fixation, Internal ; Humans ; Knee Injuries ; surgery ; Knee Joint ; surgery ; Male ; Middle Aged ; Tibia ; surgery

Objective:To explore the value of monocyte subsets and CD64 expression in the diagnosis and prognosis of sepsis.Methods:A prospective case-control study was designed. 30 septic patients and 30 non-septic patients who were admitted to the intensive care unit (ICU) of the PLA Army Characteristic Medical Center from March 2021 to March 2022 were enrolled. After 1, 3, and 5 days of ICU admission, peripheral blood samples were taken from patients. Flow cytometry was used to detect the proportion of monocyte subsets and the expression level of CD64 on the surface, and the difference of expression between patients in two group was analyzed. The risk variables for sepsis were analyzed using single-factor and multi-factor Logistic regression. The diagnostic efficacy of each risk factor for sepsis was determined using the receiver operator characteristic curve (ROC curve).Results:One day after ICU admission, the proportions of monocytes and classic monocytes in white blood cells (WBC) of septic patients were significantly lower than those of non-septic patients [proportion of monocytes to WBC: (4.13±2.03)% vs. (6.53±3.90)%, proportion of classic monocytes to WBC: 1.97 (1.43, 2.83)% vs. 3.37 (1.71, 5.98)%, both P < 0.05]. The proportion of non-classical monocytes in monocytes was significantly higher in septic patients than that in non-septic patients [(11.42±9.19)% vs. (6.57±4.23)%, P < 0.05]. The levels of CD64 expression in monocytes, classic monocytes, intermediate monocytes and non-classic monocytes were significantly higher in sepsis patients than those in non-septic patients [mean fluorescence intensity (MFI): 13.10±6.01 vs. 9.84±2.83 for monocytes, 13.58±5.98 vs. 10.03±2.84 for classic monocytes, 13.48±6.35 vs. 10.22±2.99 for intermediate monocytes, 8.21±5.52 vs. 5.79±2.67 for non-classic monocytes, all P < 0.05]. Multivariate Logistic regression research showed that CD64 in typical monocytes [odds ratio ( OR) = 1.299, 95% confidence interval (95% CI) was 1.027-1.471, P = 0.025] and the proportion of non-typical monocytes in monocytes ( OR = 1.348, 95% CI was 1.034-1.758, P = 0.027) were the independent risk factors for sepsis. ROC curve showed that the area under the ROC curve (AUC) of CD64 expression of classical monocytes, the fraction of non-classical monocytes in monocytes, and procalcitonin (PCT) in the diagnosis of sepsis was 0.871. A correlation analysis revealed a negative relationship between the acute physiology and chronic health status evaluation Ⅱ (APACHE Ⅱ) on the first, third, and fifth days following ICU admission and the expression level of CD64 in patients' classic monocytes ( r values were -0.264, -0.428 and -0.368, respectively, all P < 0.05). Conclusions:Combining the proportion of non-classical monocytes in monocytes, the level of plasma PCT, and the CD64 expression of classic monocytes in peripheral blood has good efficacy in identifying sepsis and assessing its severity.

Objective To investigate the relationship between interleukin 10 (IL-10) -592 (rs1800872) and -819 (rs1800871) promoter genetic polymorphisms and the susceptibility of antituberculosis drug-induced liver injury (ADLI). Methods A case-control study was conducted. Epidemiology survey data and peripheral blood samples were obtained from the patients. Two IL-10 gene polymorphisms (-592 A/C and 819 C/T) were genotyped with PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) in Chinese Han ADLI subjects (n=180) and sex matched by frequency matching in control subjects (n=180). Results No significant differences in genotypes of IL-10 -592 site and IL-10 -819 site between ADLI group and that of the control group were noticed (all P>0.05). The mutant alleles -592 C of IL-10 gene polymorphism was significantly higher in ADLI subjects compared to controls, and in dominant model, the frequency of CC+AC genotype was 1.62 higher among the cases than controls (all P<0.05). Significant difference in allele -819 C/T between the ADLI group and the control group were not found (P=0.190). The polymorphisms at -819 C/T and -592 A/C variants of IL-10 gene were found to be good linkage disequilibrium. The CC haplotype represent genetic risk factor (OR=1.37, 95% CI: 1.02-1.85) and CA haplotype represent genetic protect factor (OR=0.49, 95% CI: 0.34-0.70) for ADLI in the subjects. Conclusions The polymorphisms in IL-10 gene -592 A/C and -819 C/T are associated with ADLI.

OBJECTIVETo study the effects of combination of angiopoietin-1 (ANG-1) and vascular endothelial growth factor₁₆₅ (VEGF₁₆₅) gene transfer mediated by recombinant adeno-associated viral vector on the neovascularization in chronic ischemic porcine myocardium.

METHODSAn ameroid constrictor was implanted around the left circumflex coronary artery (LCX) via endoscopy. Six weeks later, coronary angiography revealed that the myocardial ischemia was established by gradual occlusion of the left circumflex coronary artery (LCX). Sixteen swine with the total occlusion or partial stenosis (> 85 %) of the LCX were divided into 4 groups (4 in each group): group I, group II and group IV (control) received direct myocardium injection of rAAV₂ VEGF₁₆₅, rAAV₂ ANG-1 or PBS alone, respectively; group III received rAAV₂ VEGF₁₆₅ and rAAV₂ ANG-1. Selective coronary angiography and ultrasonography were performed perioperatively to evaluate the cardiac function and the formation of collateral circulation. The expression of VEGF₁₆₅ and ANG-1 proteins were assessed using ELISA or Western blot. The degree of angiogenesis was assessed by use of immunohistochemical analysis.

RESULTAngiography showed that the occlusion of all LCX was completed or exceeded 95% 6 weeks after ameroid constrictor implantation, indicating the successful establishment of animal model. The expression levels of VEGF₁₆₅ in group I and III and ANG-1 in groups II and III began to increase at d7 after transfection and reached the peak at d14; then decreased gradually to the normal level after 3 months. The expression levels of VEGF₁₆₅ in group II and group IV or that of ANG-1 protein in group I and group IV had no markedly changes at different time after transfection. There were significant increase in capillary density and arteriole density and more side branch vessels formed in group III compared with other groups. Echocardiographic measurements showed that the left ventricular systolic function of animals in groups I, II and III increased significantly after gene transfection, especially in group III; but there was no changes in group IV.

CONCLUSIONMyocardial perfusion and the left ventricular systolic function are improved after rAAV₂ VEGF₁₆₅ or rAAV₂ ANG-1 transfection, which is associated with the angiogenesis in porcine model of chronic myocardial ischemia.

Adenoviridae ; genetics ; Angiopoietin-1 ; genetics ; Animals ; Collateral Circulation ; Coronary Vessels ; physiopathology ; Disease Models, Animal ; Genetic Therapy ; Genetic Vectors ; Male ; Myocardial Ischemia ; physiopathology ; therapy ; Neovascularization, Physiologic ; Swine ; Swine, Miniature ; Transfection ; Vascular Endothelial Growth Factor A ; genetics

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells (BCSCs). BCSCs were enriched by using serum-free medium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and flow cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a promising therapeutical approach for breast cancer.
Animals ; Breast Neoplasms ; genetics ; pathology ; Cell Line, Tumor ; Female ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Mice ; Neoplastic Stem Cells ; metabolism ; Receptor, Notch1 ; biosynthesis ; genetics ; Signal Transduction

OBJECTIVETo quantitatively detect the methylation of E-cadherin gene 5'-CpG islands in acute leukemia by microarray-based DNA analysis and to briefly discuss the role of microarry for detection of methylation in tumors.

METHODSBisulfite-modified DNA was used as a template for PCR amplification, resulting in conversion of unmethylated cytosine, but not methylated cytosine, into thymine within CpG islands of interest. Five sets of oligonucleotide probes were designed to fabricate a DNA microarray to detect the methylation changes of E-cadherin gene CpG islands in acute leukemia. By drawing a standard curve to assess the levels of changes in methylation detected in the examined samples.

RESULTSMicroarray assay was successfully used to quantitatively detect methylation changes of E-cadherin gene in 5 acute leukemia samples. Varying degree of methylation was detected in five regions and the hypermethylation region was the same. The result was validated by gene sequencing.

CONCLUSIONMicroarray assay may be applied as an useful tool for mapping methylation changes in multiple CpG loci and for leukemia research. It is more time-saving and labor-saving than gene sequencing and can be used to quantitatively detect changes in methylation with high throughput.

Base Sequence ; Cadherins ; genetics ; CpG Islands ; genetics ; DNA Methylation ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; methods ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Promoter Regions, Genetic