Objective: To describe the MICM (morphology, immunology, cytogenetics and molecular biology) characteristics of a case of acute myelomonocytic leukemia M 4C . Methods: The medical history data of the case of M 4C admitted to our hospital was reviewed. The results of bone marrow cell morphology, cytochemical stains, bone marrow biopsy, immunophenotype, cytogenetics, molecular test and NGS (next-generation sequencing) of the case were analyzed. Results: The bone marrow smear showed markedly active proliferation of bone marrow cells in which the myelomonocytic cells accounted for 85.6%. Cytochemical stains showed peroxidase (POX) stain partially and weakly positive; specific esterase AS-DCE partially positive; non-specific esterase α-NBE partially positive and smothered by sodium fluoride; non-specific esterase AS-DAE partially positive and smothered by sodium fluoride. Bone marrow biopsy showed hyperproliferative cells and diffused hyperplasia of blasts. Immunophenotype analysis showed that the abnormal cell population was positive for CD11B, CD64, CD56, cMPO, CD33, CD41, CD61, CD38 and CD58, but negative for CD13, CD34, CD117, CD7, CD123, HLA-DR, CD10, CD19, CD20, CD2, CD14, CD235, CD15, CD303, CD304, CD25, cCD79a, cCD3, cCD22, CD1a and TDT. Cytogenetic analysis showed 47, XY, t(9;11) (p22;q23),+mar. The molecular test for leukemia showed MLLT3/KMT2A gene rearrangement. NGS showed NRAS and TET2 mutation. The case was finally diagnosed as AML (acute myelomonocytic leukemia) M 4C with t(9;11)(p22;q23), MLLT3-KMT2A. Conclusion Leukemia M 4C may show the characteristics of both granulocytes and monocytes with complex morphological features. The combined examination of MICM should be necessary for the diagnosis of M 4C with great significance.

Objective To investigate the efficacy and safety of the affinity adsorption material for removal of intes-tinal endotoxin in the blood. Metheds To establish a canine model of intestine-derived endotoxemia by ligation and perfo-ration of the appendix. Hemoperfusion was performed to treat the endotoxemia in model dogs. In the experiment,we moni-tored vital signs and detect the content of endotoxin and blood physiological and biochemical indexes at different time points. Results The endotoxin content was(0.49 ± 0.22),(0.034 ± 0.00)Eu/mL after hemoperfusion for 1 and 2 hours,compared with that before the beginning of perfusion(7.25 ± 1.18)Eu/mL,showing a significant difference(P <0.05). After treatment for 2 hours,the average clearance rate was 99.52%. White blood cells,red blood cells,hemoglo-bin,platelets,total protein,albumin,globulin,alanine aminotransferase,aspartate aminotransferase,urea and creatinine index were significantly decreased after perfusion(P < 0.05). The index values were in a normal range and vital signs were stable during the perfusion. Conclusions The affinity adsorption material developed by our team can effectively re-move endotoxin in the blood of experimental dogs and has good blood compatibility.

Objective To investigate the effectiveness of the affinity adsorption material developed by our team for the specific removal of exogenous endotoxin in the blood circulation. Methods Fifteen beagle dogs were intravenously injected with endotoxin to establish a dog model of endotoxemia, and then they were randomly divided into the treatment group(n=10)and the control group(n=5). The treatment group received an extracorporeal perfusion to remove the endotoxin using the self-made disposable hemoperfusion device,while the control group using routine perfusion device. The levels of endotoxin, tumor necrosis factor α(TNF-α), interleukin 1β(IL-1β), interleukin 6(IL-6)and interleukin 8 (IL-8)in the blood of the dogs were measured at the beginning and 120 min after hemoperfusion for 120 minutes. The vital signs of the dogs were monitored during the hemoperfusion. Results After successful establishment of the endotoxemia model,the level of endotoxin at the beginning of hemoperfusion in the treatment group and control group was 118.63 ± 27.98 EU/mL and 117.16 ± 22.95 EU/mL,respectively. After hemoperfusion for 120 min,it was 0.039 ± 0.01 EU/mL and 131.98 ± 7.01 EU/mL, showing a significant difference(P﹤0.05). The clearance rate of hemoperfusion in the treatment group was 94.07%. At the beginning of hemoperfusion, the levels of TNF-α, IL-1β, IL-6 and IL-8 in the treatment group were 1.53 ± 0.27 ng/mL,12.82 ± 1.66 ng/mL,54.77 ± 3.98 ng/mL and 0.25 ± 0.32 ng/mL, and the levels in the control group were 1.53 ± 0.06 ng/mL,13.05 ± 0.18 ng/mL,54.58 ± 0.19 ng/mL and 0.28 ± 0.06 ng/mL, respectively. After hemoperfusion for 120 min, the levels of TNF-α, IL-1β, IL-6 and IL-8 in the treatment group were 0.13 ± 0.06 ng/mL, 0.70 ± 0.36 ng/mL, 1.62 ± 0.80 ng/mL and 0.01 ± 0.00 ng/mL, respectively, and as for the control group,the levels were 2.26 ± 0.15 ng/mL,15.12 ± 0.18 ng/mL,62.54 ± 0.93 ng/mL and 0.73 ± 0.93 ng/mL. There were significant differences between the beginning and after perfusion for 120 min in those two groups(P< 0.05). Conclusions This affinity adsorption material can effectively remove endotoxin and the inflammatory mediators in the blood of experimental dogs,with a clearance rate of 94.07%.

Objective To investigate the tumor/ non-tumor (T/ NT) ratio during 99 Tcm-hydrazinon-icotinamide(HYNIC)-( poly-( ethylene glycol), PEG) 4-Glu( cyclo ( Arg-Gly-Asp-D-Phe-Lys ( PEG4 ))) 2 ( 99 Tcm-3PRGD2 ) SPECT/ CT imaging and clinical pathological features of breast cancer. Methods Forty-five female patients (age range: 39-76 (53.0±9.5) years) with suspected breast malignant nodules or mas-ses from October 2016 to June 2017 were prospectively enrolled. Patients underwent 99 Tcm-3PRGD2 SPECT/CT imaging before breast puncture and surgery. All subjects had pathological results, and estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2(HER-2), Ki-67 and mi-crovessel density ( MVD) were obtained by immunohistochemistry and fluorescence in situ hybridization(FISH). Two-sample t test and Kruskal-Wallis H test were used to analyze the data. Results Invasive duc-tal carcinoma was pathologically confirmed in 35 of 45 patients. There were 7 patients in stage Ⅰ, 11 pa-tients in stage ⅡA and 17 patients in stage ⅡB. The Luminal A subtype, Luminal B subtype, ERBB2+subtype, Basal-like subtype were found in 6, 9, 9 and 11 patients, respectively. The T/ NT ratio was signifi-cantly higher in the stageⅡB patients than that in stageⅠ+ⅡA patients (4.54±1.46 vs 3.32±1.72, t= -2.24, P<0.05). Patients with ERBB2+ subtype had higher T/ NT ratio compared to patients with Basal-like sub-type: 5.80(3.90, 6.70) vs 2.80(2.20,3.50), H= 11.06, χ2 = 15.31, both P<0.05. Besides, the T/ NT ra-tios in the HER-2 positive group and lymphatic metastasis group were significantly higher than those in the HER-2 negative group and group without lymph node metastasis (t values: -3.99, -2.51, both P<0.05). MVD of HER-2 positive group was higher than that of HER-2 negative group (t= 7.13, P<0.01). Conclu-sion The T/ NT ratio during 99 Tcm-3PRGD2 SPECT/ CT imaging has relations with TNM staging, lymph node infiltration and HER-2 in breast cancer.

BACKGROUND: Endoplasmic reticulum (ER) stress has been proved to be related to the occurrence of diabetes, dilated cardiomyopathy and neurodegenerative diseases. Indeed, it is closely associated with osteoarthritis. OBJECTIVE: To explore the effect of ER stress on the chondrocyte viability as well as the occurrence and development of osteoarthritis in rats. METHODS: Rat chondrocytes were isolated and cultured, and the ER stress in the rat chondrocytes was by 10 mg/L tunicamycin. The expression levels of ER stress markers C/EBP-homologous protein and 78 kDa glucose-regulated protein were detected by western blot assay, and the proliferation and apoptosis of chondrocytes were detected by cell counting kit-8 assay and AnnexinV-FITC flow cytometry, respectively. In the in vivo experiment, 15 Sprague-Dawley rats were selected and subjected to anterior cruciate ligament transection and medial meniscectomy to establish an animal model of osteoarthritis. Tunicamycin, tauroursodeoxycholic acid and PBS (blank control group) were respectively injected into the articular cavity, and then the progression of osteoarthritis was assessed by hematoxylin-eosin staining at 4 weeks after treatment. RESULTS AND CONCLUSION: After addition of tunicamycin, the expression levels of C/EBP-homologous protein and 78 kDa glucose-regulated protein were significantly upregulated, the viability of chondrocytes was decreased gradually, while the apoptotic rate was increased significantly. Results from gross observation and hematoxylin-eosin staining suggested that tunicamycin promoted the progression of osteoarthritis and tauroursodeoxycholic acid delayed the deterioration of cartilage in the rats. These findings indicate that ER stress results in the decreased chondrocyte viability and increased apoptosis, which may be an important pathogenesis of osteoarthritis. Additionally, tauroursodeoxycholic acid can effectively alleviate osteoarthritis induced by ER stress.

Objective:To investigate whether endogenous hydrogen sulfide (H2 S)was involved in the pathogenesis of osteoarthritis (OA)and its underlying mechanism,to detect H2 S and its synthases ex-pression in knee cartilage in patients diagnosed with different severity of OA,and to explore the transcrip-tion and expression of gene MMP-13 in chondrocytes treated with IL-1βor H2S.Methods:Synovial fluids of the in-patients with different severity of OA hospitalized in Peking University First Hospital were collected for measurement of H2 S content using methylene blue assay.Articular cartilages of the patients who underwent knee arthroplasty were collected for the cell culture of relatively normal chondrocytes.The chondrocytes were cultured to the P3 generation and H2 S molecular probes were used for detection of endogenous H2 S generation in the chondrocytes.Immunocytochemistry was used to detect the localization of H2 S synthases including cystathionine β-synthase (CBS),cystathionine-γ-lyase (CSE),and mercap-topyruvate sulfurtransferase (MPST)in OA chondrocytes.Western blot was used to quantify the protein expressions of CSE,MPST,and CBS in cartilage tissues of the patients who were diagnosed with OA and underwent knee arthroplasty.The relatively normal human chondrocytes were cultured to passage 3 and then divided into 4 groups for different treatments:(1 )the normal control group,no reagent was added;(2)the IL-1βgroup,5 μg/L of IL-1βwas added;(3)the IL-1β+H2S group,200 μmol/L of NaHS was added 30 min before adding 5 μg/L of IL-1β;(4)the H2 S group,200 μmol/L of NaHS was added. The transcription and expression of gene MMP-13 in chondrocytes of each group were determined with Real-time PCR and Western blot,respectively.And the total NF-κB p65 and phosphorylated NF-κB p65 in chondrocytes were detected with Western blot.Results:The content of H2 S in the synovial fluid of degenerative knee was (14.3 ±3.3)μmol/L.Expressions of endogenous H2 S and its synthases including CBS,CSE and MPST were present in the cytoplasm of chondrocytes.CSE protein expression in Grade 3 (defined by outerbridge grading)cartilage tissues was significantly increased as compared with that of Grade 1 cartilage tissues (1.67 ±0.09 vs.1.26 ±0.11,P<0.05).However,no significant difference of CBS or MPST expression among the different groups was observed.The expression of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (1 .87 ±0.67 vs.0.22 ± 0.10,P<0.05 ),and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (0.55 ±0.11 vs.1.87 ±0.67,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The transcription of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (31.40 ±0.31 vs.1.00 ±0.00,P<0.05), and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (24.41 ± 1.28 vs.31.40 ±0.31,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The total NF-κB p65 in the IL-1βgroup was significantly higher than that in the normal chondrocytes (2.13 ±0.08 vs.0.73 ±0.08,P<0.05),and that in the IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (1 .24 ±0.13 vs.2.13 ±0.08,P<0.05 ),and that in the H2 S group had no significant difference compared with that in the normal control group.The phosphorylated NF-κB p65 in IL-1βgroup was significantly higher than that in the normal chondrocytes (1.30 ±0.13 vs.0.19 ±0.04,P<0.05),and that in IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (0.92 ±0.26 vs.1.30 ±0.13,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.Conclusion:H2 S affected the cartilage degeneration by partly inhibiting the degradation of extracellular matrix.

Objective To explore the experience of diagnosis and surgical treatment of carotid body tumor.Methods A retrospective analysis between November 2008 and November 2015 was proceeded,the clinical data of surgical treatment for 81 patients with carotid body tumor was collected,to analyze data by SPSS19.0,and summarize the diagnosis of carotid body tumor,choice of operation methods and curative effect and complications prevention.Results Seventy-four cases underwent surgery treatment:tumors of 52 cases were simply stripped,tumors of 13 cases were resected combined with ligation of external carotid artery.Tumors of 7 cases were resected with internal and external carotid artery ligation,3 cases of whom underwent artificial blood vessel internal carotid artery end to end anastomosis.Postoperative death in 1 case of acute myocardial infarction,complicated with cerebral infarction in 2 cases,6 cases of injury of cranial nerve relieved after symptomatic treatment.No hemiplegia,aphasia and other serious complications.Tumor size and the surgery time correlation analysis:the correlation coefficient was 0.226,no significant correlation.Conclusions CTA is the most commonly used method of preoperative examination.Surgical resection is an effective method in treatment of carotid body tumor.Prevention injury of carotid artery cr internal carotid or common carotid artery and their reconstruction is the key to a successful operation.Sufficient preoperative assessment,select the appropriate operation method,intraoperative careful performance can ensure the cerebral perfusion,is the key to prevent and reduce the complications.

Objective To retrospectively study the risk factors of aortic arch calcificationand its influence on the survival prognosis of maintenance peritoneal dialysis patients. Methods One hundred seventy-seven cases of maintenance peritoneal dialysis patients were enrolled, including 66 cases of aortic arch calcification cases. Their general dialysis data were collected for the evaluation of dialysis adequacy and residual renal function, and their chest X-rays were recorded to assess the degree of aortic arch calcification. The two variables Logistics regression was used to analyze independent risk factors of aortic arch calcification; Kaplan-Meier analysis was used to analyze the influence on prognosis of dialysis patients; and multivariate COX regression was employed to analyze independent risk factors of death in dialysis patients. Results Among the 177 selected cases of peritoneal dialysis patients, 66 cases (37.29%) presented with aortic arch calcification. Elevated serum phosphorus was an independent risk factor of aortic arch calcification (OR=54.69 ,95%CI:10.01-298.65, P<0.01). The probability of survival in patients with mild and moderate (severe) calcification of aortic arch was less than those without calcification. Moderate (severe) calcification of aortic arch was the independent risk factor of all-cause mortality and cardiovascular disease mortality, whose hazard ratios in patients with calcification were 3.779 times and 5.636 times of those in patients without calcification respectively. Conclusions Hyperphosphatemia is an independent risk factor promoting the development of calcification of aortic arch. The probability of survival in patients with mild and moderate (severe) calcification of aortic arch is less than those without calcification; moderate (severe) calcification of aortic arch is the independent risk factor of all-cause mortality and cardiovascular disease mortality.

Objective:To investigate the efficacy of single time intra-articular different concentration of allogeneic bone marrow mesenchymal stem cells ( BM-MSCs ) injection in the treatment of Sprague-Dawley ( SD) rat model of osteoarthritis ( OA) .Methods: In the study, 32 SD rats were equally ran-domized into 4 groups:control group, high concentration group (1 ×107/mL BM-MSCs), low concentra-tion group (5 ×106/mL BM-MSCs) and high vs.low concentration group.The two knees of each rat were set up to a pair.The induction of OA was performed surgically randomly at one side in model group, and bilaterally in the other groups, which were through anterior cruciate ligament transaction ( ACLT) and medial meniscus excising.After the operation, the SD rats were allowed free movement.Four weeks later, different concentrations of allogeneic BM-MSCs isolated from the SD rats, expanded in vitro and suspended in phosphate buffered solution( PBS) were delivered in the articular cavity of both knees;PBS was used as the control.After injection, we excised the femoral nerve and sciatic nerve to disuse the low limb.The cartilage histological sections of knees were scored by Mankin scoring system to assess the se-verity of the pathology.mRNA of collagen Ⅱwas detected by real time polymerase chain reaction ( RT-PCR) .eGFP was detected by fluorescence microscope.Assessments were carried out 4 weeks after the operation in model group, and 3 weeks after injection in the other groups.Results:Mankin scores of the BM-MSCs side and control side were 6.60 ±0.40 vs.10.00 ±0.32 in low concentration group ( P<0.05), and 5.40 ±0.51 vs.9.60 ±0.51 in high concentration group (P<0.05).Mankin scores of high sv.low concentration group were 6.40 ±0.51 vs.7.60 ±0.75 (P>0.05).mRNA expression of collagen Ⅱ of the BM-MSCs side in low concentration group was 106%±1%in contrast to the control side.As in high concentration group it was 108%±1%, and 102%±1%in high vs. low concentra-tion group.Labeled BM-MSCs were detected unexpectedly in the synovial membrane but not in cartilage tissue three weeks from injection.Conclusion:BM-MSCs could promote cartilage repair and inhibit OA progression through a trophic mechanism.There was no difference between the two concentrations.