Objective:To observe and analyze the clinical features and prognosis of proliferative diabetic retinopathy (PDR) with chronic myeloid leukemia.Methods:A retrospective case series study. From May 2011 to December 2020, 5 patients (10 eyes) were included in this study in Eye-ENT Hospital of Fudan University. Basic information about the patient's age, gender, diabetes history and CML history were collected. The endocrine and hematological indexes of all patients were evaluated. All the patients were undertaken visual acuity, intraocular pressure, slit lamp and fundus examination and other examinations to observe the eye conditions. Ophthalmic treatments included panretinal laser photocoagulation, intravitreal injection of anti-vascular endothelial growth factor, vitrectomy. During the follow up period from 5 months to 6 years, prognosis was observed at each office visit. During the follow up period, patients' vision, intraocular pressure, anterior segment and retinal status were observed.Results:There were 4 males and a female in 5 patients. The ages were from 27 to 49 years, with the mean age of 39 years. All patients were bilateral. All patients suffered type 2 diabetes for 3 months to 13 years. Four of them were diagnosed as chronic myeloid leukemia before visiting to ophthalmologists, while the other visited to ophthalmology first due to poor vision. The initial visual acuity ranged from light perception to 0.4 and 6 eyes were less than 0.1. In addition to the typical manifestations of diabetic retinopathy, such as venous tortuous dilation, exudation, microaneurysm and neovascularization, patients also presented with Roth spot as leukemic fundus manifestations. All eyes developed to PDR stage. Abnormal thickening of the neovascular membranes may occur in the lower part of the retina, with secondary traction retinal detachment. All the eyes were treated with pan retinal photocoagulation and 9 eyes underwent pars plana vitrectomy. After treatment, retina of 8 eyes kept flat. The best corrected visual acuity ranged from no light perception to 1.0, and only 4 eyes reached more than 0.2. Unfortunately, one eye lost vision because of secondary neovascular glaucoma.Conclusions:PDR patients with CMLof fundus not only have venous tortuous dilation, exudation, microaneurysm and neovascularization, also present with Roth spot as leukemic fundus manifestations. Diabetic retinopathy combined with CML could progress rapidly, and its aggravating complications such as hyperplastic membrane, vitreous hemorrhage and traction retinal detachment may result in poor visual prognosis. Early screening and treatment can help improve the prognosis of patients.

Objective:To develop a biomimetic nanoparticle probe of hematoporphyrin monomethyl ether (HMME) coated with breast cancer cell membrane, to observe its ability to target homologous breast cancer cells in vitro, and to investigate its effect of enhanced photoacoustic imaging and sonodynamic therapy (SDT) for breast cancer in vitro.Methods:The cell membrane of breast cancer 4T1 was extracted by chemical cleavage and repeated freezing and thawing. Then the HMME-coated polylactic acid-glycolic acid copolymer biomimetic nanoparticle was prepared by double emulsification and extrusion. The basic characteristics of nanoparticles were detected. The target ability of nanoparticles to homologous breast cancer cells and the enhancement of photoacoustic imaging were observed in vitro. Singlet oxygen sensor green (SOSG) was used to verify the reactive oxygen species (ROS) production of nanoparticles, and its SDT effect on breast cancer cells was evaluated by CCK8 cytotoxicity assay.Results:The size of the prepared CHP-NPs was uniform, the morphology was spherical "core-shell structure" , the particle size was (275.23±8.25)nm, and the surface potential was (-18.43±0.45)mV. It was observed that CHP-NPs could target homologous 4T1 cells under laser confocal microscopy. In vitro photoacoustic imaging experiments show that the photoacoustic signal of nanoparticles increases with the increase of its concentration. According to SOSG probe detection, CHP-NPs could produce ROS under ultrasonic irradiation.When CHP-NPs was incubated with 4T1 cells alone and no ultrasonic irradiation was used, the cell survival rate was not significantly affected. When the concentration was 0.6 mg/ml, the cell survival rate was still 95%. After ultrasonic irradiation, CCK8 experiment showed that the CHP-NPs had a significant SDT effect on breast cancer cells.Conclusions:The biomimetic nanomolecular probe of breast cancer cell membrane is successfully prepared. The probe has good ability to target homologous tumor, and can significantly enhance tumor photoacoustic imaging and SDT effect.

OBJECTIVETo study the effect of alternatively activated macrophages /mononuclear phagocytes(MNP) on breast cancer cells and explore the mechanisms for the action of tumor-associated macrophages in breast cancer.

METHODSHuman peripheral blood monocytes were isolated and cultured in vitro and divided into 3 groups, namely classically activated monocytes (CAM) which were induced by lipopolysaccharide, alternatively activated monocytes (AAM) induce by IL-4, and control cells treated with the culture medium only. After cell culture for 48-72 h, the mRNA of tumor necrosis factor-alpha (TNF-alpha), alternative monocytes activation- associated CC-chemokine 1 (AMAC-1), and beta-actin of the 3 groups were extracted for RT-PCR, or the cells were cocultured with breast cancer cell line SKBR3, or seeded in chicken chorioallantoic membrane along with SKBR3.

RESULTSTNF-alpha mRNA was significantly increased in CAM, and AMAC-1 was highly expressed in AAM. The coculture experiments showed that CAM exhibited obvious inhibitory effect on SKBR3 cells after a 3-day culture whereas AAM significantly promoted the growth of SKBR3 cells after a 5-day culture. In chicken on chorioallantoic membrane experiment, the macrophages promoted tumor angiogenesis and AAM showed the most obvious effect.

CONCLUSIONIL-4 induces high expression of AMAC-1, a molecular marker of AAM, in the macrophages, and AAM can promote the growth of SKBR3 cells and tumor angiogenesis.

Animals ; Breast Neoplasms ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Chemokines, CC ; metabolism ; Chick Embryo ; Coculture Techniques ; Humans ; Interleukin-4 ; metabolism ; Macrophage Activation ; Phagocytes ; immunology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the role of protein kinase C-delta (PKC-delta) in hyperthermia-induced apoptosis in human tongue squamous cell carcinoma Tca8113 cells.

METHODSTca8113 cells were treated at 43 degrees C in a heating water bath for 0, 40, 80, 120 min after pretreatment with Rottlerin, a specific inhibitor of PKC-delta, and equal volume dimethyl sulfoxide (DMSO) for 30 min, respectively. The cells were stained by propidium iodide (PI) and Rhodamine 123 to analysis apoptotic rate and the changes of mitochondrial transmembrane potential by flow cytometry (FCM). The total proteins were extracted for Western blotting analysis of activation and proteolysis of PKC-delta, and for colorimetric assay of relative activity of Caspase-3.

RESULTSHyperthermia could induce proteolysis and activation of PKC-delta, and this was attenuated by Rottlerin. Apoptotic rate, decreasing of mitochondrial transmembrane potential and activity of Caspase-3 which being induced by hyperthermia in Tca8113 cells were inhibited by PKC-delta specific inhibitor Rottlerin. There were significantly statistical differences in apoptosis rates, mitochondrial transmembrane potential and activity of Caspase-3 between Rottlerin- and non-Rottlerin-pretreated cells after hyperthermia for 40, 80, 120 min (P < 0.01).

CONCLUSIONActivated PKC-delta may facilitate hyperthermia-induced apoptosis in Tca8113 cells, and may be one of the mechanisms of apoptosis induced by hyperthermia.

Acetophenones ; Apoptosis ; Benzopyrans ; Carcinoma, Squamous Cell ; Humans ; Protein Kinase C ; Protein Kinase C-delta

OBJECTIVETo observe the incidence of heparin-induced thrombocytopenia (HIT) in patients received unfractionated heparin (UFH) treatment, and explore the feasibility of monitoring HIT by platelet counts, as well as the significance of HIT-antibody test in HIT diagnosis.

METHODS145 patients received UFH treatment in Vascular Surgery Department were studied. Before and after the UFH treatment, platelet counts, HIT-antibody ELISA test and heparin-induced platelet aggregation (HIPA) were tested.

RESULTSAmong the 145 patients, thrombocytopenia occurred in 40 (27.6%) cases, HIT-antibody ELISA test positive in 59 (40.7%) cases, HIPA test positive in 26 (17.9%) cases. The HIT was diagnosed in 24 (16.5%) cases, and heparin-induced thrombocytopenia and thrombosis (HITTS) occurred in 5 (3.4% in all cases, and 20.8% in HIT patients). In HIT patients, 15 patients (62.5%) were thrombocytopenia, HIT-antibody positive and HIPA test positive. Platelet counts in all of the 24 patients recovered to normal or level before UFH treatment in 3-6 days after heparin withdrawal therapy.

CONCLUSIONHIT can be early diagnosed by monitoring platelet counts, HIT-antibody ELISA test and HIPA test. Withdrawal of heparin therapy in time and use of alternative anticoagulant, HITTS rate might be expected to decline further.

Adult ; Aged ; Anticoagulants ; adverse effects ; Enzyme-Linked Immunosorbent Assay ; Female ; Heparin ; adverse effects ; Humans ; Male ; Middle Aged ; Platelet Aggregation ; Platelet Count ; Thrombocytopenia ; chemically induced ; diagnosis

Objective To introduce the application experience of a new steel bar used in minimally invasive surgery for pectus carinatum.Methods From January to October 2018, Cardiothoracic Surgery Department of Shanghai Xinhua Hospital performed a minimally invasive surgery for 25 cases of patients with pectus carinatum used a new type of steel bar.All 25 pa-tients were male, aged 10 -17 years, with an average age of(13.80 ±1.66)years.The application experience of the new bar in pectus carinatum minimally invasive surgery was summarized .Results All operations were successfully completed .The op-eration time was 35-100 min, averaged(73.44 ±17.49)min, postoperative hospital stay was 3 -6 days, averaged(3.68 ± 0.85)days.Postoperative complications included 5 cases of pneumothorax(the lung compression was about 2％ -10％, not necessary for surgical intervention).One case occured wound healing delay 1 month after operation, and healed after no surger-cal treatment.The other patients recovered smoothly.Conclusion The new steel bar is convenient to use, greatly reduces the difficulty of the pectus carinatum surgery procedure , also reduced surgical trauma and complications , has a good application prospect.

OBJECTIVETo sort CD133(+) subset cells in human gastric cancer (GC) and to identify their tumor initiating cell-like properties.

METHODSThe tissues of GC and normal tissues adjacent to GC were obtained from 50 patients. Samples were stained for CD133 by immunohistochemistry. Likewise, assessments of CD133 were undertaken by Western blot. Flow cytometry was used to determine the proportion of CD133(+) cells in four GC cell lines therein the KATO-III was sorted by magnetic activated cell sorting (MACS) method. The growing characteristics and the tumorigenic ability of CD133(+) cells were evaluated in vitro and in vivo. Meanwhile, the growth of single cells in suspension culture was observed and expression of stem cell-specific marker were determined using reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe expression of CD133 was demonstrated on the cell membranes in the mucosa and submucosa of primary GC, which were higher than those in the normal gastric tissues adjacent to cancer (P<0.05). Four GC cell lines including KATO-III, SGC-7901, AGS and MKN-45 were found to contain (28 ± 2)%, (17 ± 2)%, (6 ± 2)%, and (4 ± 2)% of CD133(+) cells respectively. In addition, the purity of CD133(+) cells isolated from KATO-III by MACS was (91 ± 3)% and up to(95 ± 2)% after 1-week culture. CCK-8 detection showed that population doubling time of the CD133(+) cells was (21 ± 3)h, significantly shorter than that of the CD133(-) cells[(40 ± 8)h, P<0.05]. Notably, there was a remarkable difference of tumor formation rate between CD133(+) cells (100%), non-sorted cells (80%), and CD133(-) cells(0). The average mass and volume of tumor in group of CD133(+) cells was larger and heavier than those in non-sorted cells (P<0.05, P<0.05). Furthermore, the single cell proliferated well, formed the big sphere and semi-quantitative RT-PCR showed expression of stem cell markers such as Oct-4, Nanog, Sox-2, Musashi-1 and EGFR.

CONCLUSIONSCD133 protein expression in primary lesions is higher than those in the normal gastric tissues. CD133(+) subset cells can be isolated, purified, and amplified in human GC, and possess some properties including the ability of self-renewal, proliferation, and higher tumorigenic ability in vivo and can express some stem cell markers.

AC133 Antigen ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antigens, CD ; metabolism ; Cell Line, Tumor ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Middle Aged ; Neoplastic Stem Cells ; metabolism ; Peptides ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Xenograft Model Antitumor Assays

OBJECTIVETo study the feasibility of vector-mediated RNA interference for HER-2-positive breast cancer therapy.

METHODSA plasmid vector capable of mediating HER-2 RNA interference was constructed, and HER-2-positive breast cancer cell line SKBR-3 was transfected with this constructed vector. The expression of HER-2 mRNA and protein was analyzed by RT-PCR and Western blotting, and the growth and apoptosis of SKBR-3 cells was analyzed after transfection.

RESULTSThe expressions of HER-2 mRNA and HER-2 protein was downregulated in response to vector-mediated HER-2 RNA interference, which also resulted in tumor cell growth inhibition and increased number apoptotic cells.

CONCLUSIONHER-2 is a good target for RNA interference and RNA interference targeting HER-2 can lead to HER-2 breast cancer cell apoptosis and growth inhibition.

Apoptosis ; Blotting, Western ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Genetic Vectors ; Humans ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Receptor, ErbB-2 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo investigate the changes in proliferation, invasiveness, clone sphere formation and chemosensitivity of human gastric cancer cell lines of KATO-III CD133(+) cells transfected with small interfering RNA (siRNA) against CD133 gene.

METHODSCD133(+) cells of KATO-III cell lines were isolated by magnetic activated cell sorting (MACS). CD133 siRNA was designed and synthesized, and then transfected into KATO-III CD133(+) cells. Cell fluorescence counting under confocal laser scanning microscope was used to determine the transfection efficiency after transfection with the CD133 FITC-siRNA. The knock-down effect of the CD133 gene and expression of epithelial-mesenchymal transition (EMT)-related factors were detected by RT-PCR and Western blotting. Cell counting kit-8 assay (CCK-8), transwell chamber and colony sphere forming assay were performed to measure the variation of cell proliferative, invasive, colony formation viability and chemosensitivity to 5-FU after the above-mentioned treatment.

RESULTSThe transfection efficiency was (87.7±8.1)%. The CD133 mRNA and protein expression levels in the interference group were lower than those in negative control group. Twenty-four, 48 and 72 hours after transfection, cells proliferation activity was significantly inhibited in the interference group compared with negative control group, (all P<0.01). Seventy-two hours after transfection, compared with negative control group, cells proliferation activity was reduced by (52.1±8.0)%. The invasive cell number reduced (41.7±6.0 vs. 130.3±11.0, P<0.05) and clone formation rate decreased significantly [(24.3±4.3)% vs. (45.1±6.4)%, P<0.01] in the interference group. EMT-related gene E-cadherin protein expression increased, while the Snail and N-cadherin protein expression reduced in the interference group (all P<0.01). The cells sensitivity to 5-FU was significantly enhanced in the interference group, and the cell inhibition rate of 5-Fu was (62.4±3.3)%, higher than that in negative control group [(21.5±2.2)%, P<0.01].

CONCLUSIONSThe expression of CD133 gene plays an important role in cell proliferation, invasiveness, colony formation and resistance to chemotherapy of KATO-III CD133(+) gastric cancer cells. It suggests that CD133 can be used as one of surface markers for detection of gastric cancer stem cells. Inhibition of CD133 expression may be a promising way for gastric cancer biotherapy.

AC133 Antigen ; Antigens, CD ; genetics ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Fluorouracil ; pharmacology ; Glycoproteins ; genetics ; metabolism ; Humans ; Peptides ; genetics ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection

OBJECTIVETo examine the association between CD133 expression and invasion of gastric cancer, and to elucidate whether CD133 can promote the invasion and metastasis of gastric cancer via epithelial-mesenchymal transition (EMT).

METHODSThe CD133(+) and CD133(-) KATO-III( cells were sorted by magnetic activated cell sorting (MACS). The invasion ability was detected by Transwell method. RT-PCR and Western blot were used to detect the expression of EMT-related factors in KATO-III( cells before and after CD133 was knocked out by siRNA method. The expressions of CD133 and EMT-related proteins of cancer and adjacent normal tissues in 50 patients with gastric cancer were detected by Western blot, and correlations among protein expressions were also analyzed.

RESULTSAs compared to CD133(-) cells, the number of broken-membrane cells was significantly higher (67.7±10.5 vs. 13.3±6.8, P=0.001) and the invasion ability was stronger (P<0.05) in CD133(+) cells, while the mRNA expression levels of Snail and N-cadherin were significantly higher in CD133(+) cells (0.311±0.015 vs. 0.223±0.016, P=0.040; 0.581±0.020 vs. 0.270±0.018,P=0.004), and the protein expression levels of Snail and N-cadherin were significantly higher in CD133(+) cells as well (0.513±0.015 vs. 0.179±0.023, P=0.030; 0.538±0.028 vs. 0.202±0.032, P=0.020), but E-cadherin mRNA and protein levels were significantly lower in CD133(+) cells (0.231±0.009 vs. 0.460±0.015, P=0.040; 0.426±0.030 vs. 0.748±0.027, P=0.040). After CD133 knock-out, the expressions of Snail and N-cadherin were down-regulated (P<0.05) and the expression of E-cadherin was up-regulated (P<0.05). As compared to normal mucosal tissues, the protein expression levels of Snail, N-cadherin and CD133 in gastric cancer tissues were significantly higher(0.635±0.119 vs. 0.485±0.116, P=0.029; 0.599±0.114 vs. 0.259±0.108, P=0.020; 0.754±0.154 vs. 0.329±0.134, P=0.001), while the protein expression of E-cadherin in gastric cancer tissues was lower (0.378±0.123 vs. 0.752±0.156, P=0.003). The protein expressions of Snail and N-cadherin were positively correlated with CD133 expression (r=0.278, P=0.048; r=0.406, P=0.003) and the protein expression of E-cadherin was negatively correlated with CD133 expression (r=-0.504, P=0.000).

CONCLUSIONCD133(+) cells in primary lesion of gastric cancer have relatively higher invasion ability, which may promote the metastasis of gastric cancer via up-regulation of EMT-related factors.

AC133 Antigen ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Peptides ; metabolism ; Stomach Neoplasms ; metabolism ; pathology