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Immunoscenescence plays a key role in the initiation and development of tumors. Furthermore, immunoscenescence also impacts drug delivery and cancer therapeutic efficacy. To reduce the impact of immunosenescence on anti-tumor therapy, this experimental plan aimed to use neutrophils with tumor tropism properties to deliver sialic acid (SA)-modified liposomes into the tumor, kill tumor cells via SA-mediated photochemotherapy, enhance infiltration of neutrophils into the tumor, induce immunogenic death of tumor cells with chemotherapy, enhance infiltration of CD8+ T cells into the tumor-draining lymph nodes and tumors of immunosenescent mice, and achieve SA-mediated photochemotherapy. We found that CD8+ T cell and neutrophil levels in 16-month-old mice were significantly lower than those in 2- and 8-month-old mice; 16-month-old mice exhibited immunosenescence. The anti-tumor efficacy of SA-mediated non-photochemotherapy declined in 16-month-old mice, and tumors recurred after scabbing. SA-mediated photochemotherapy enhanced tumor infiltration by CD8+ T cells and neutrophils, induced crusting and regression of tumors in 8-month-old mice, inhibited metastasis and recurrence of tumors and eliminated the immunosenescence-induced decline in antitumor therapeutic efficacy in 16-month-old mice via the light-heat-chemical-immunity conversion.
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Objective:To assess the value of CD4 + and CD8 + T-lymphocyte counts for the diagnostic classification and prognosis of coronavirus disease 2019 (COVID-19). Methods:A total of 95 COVID-19 adult patients admitted to Shanghai Public Health Clinical Center, Fudan University from January to March 2020 were recruited. The CD4 + and CD8 + T-lymphocyte counts among ordinary, severe and critical patients, as well among the cured, improved, unimproved and death patients were compared. The area under receiver operating characteristic curve (AUROC) was used to evaluate the value of CD4 + and CD8 + T-lymphocyte counts for the clinical diagnosis and prognosis of COVID-19. The comparison among groups was performed by Mann-Whitney U test. Results:A total of 95 COVID-19 cases including 68 common, 11 severe and 16 critical cases were enrolled. The counts of CD4 + and CD8 + T-lymphocyte of patients in common, severe and critical groups were 419 (309, 612), 267 (212, 540), 141 (77, 201)/μL, and 238 (153, 375), 128 (96, 172), 92 (51, 144)/μL, respectively, with significant differences ( Z=24.322 and 15.956, respectively, both P<0.01). The counts of CD4 + and CD8 + T-lymphocyte of the death, unimproved, improved, and cured patients were 149 (143, 349), 315 (116, 414), 344 (294, 426), 745 (611, 966)/μL, and 106 (43, 501), 176(67, 279), 194(188, 432), 429(276, 564)/μL, respectively, with significant differences ( Z=36.083 and 16.658, respectively, both P<0.01). The optimal cut-off point of CD4 + T-lymphocyte counts was 237/μL for critical COVID-19 with AUROC 0.911 (95% confidence interval ( CI) 0.833-0.989, P<0.01), with the sensitivity of 86.1% and specificity of 87.5%. For predicting severe and critical cases, the optimal cut-off point of CD4 + T-lymphocyte counts was 405/μL with AUROC 0.863 (95% CI 0.727-0.999, P=0.001), with the sensitivity of 78.6% and specificity of 74.6%. Conclusions:The conditions of patients with COVID-19 are aggravated with CD4 + and CD8 + T-lymphocyte counts decreasing. CD4 + T-lymphocyte counts may be an indicator for diagnostic classification of COVID-19 and prognostic indicator for severe and critical patients.
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Objective To evaluate the effect and mechanism of bone morrow MSCs transplantation in swine with AMI by cell biology and molecular imaging methods including PET/CT, SPECT, and MRI. Methods Twenty?four Chinese mini?swine ( ( 25 ± 5 ) kg ) were randomly divided into 2 groups: MSCs group ( n=12) and control group ( n=12) . Myocardial infarction was induced in swine hearts by occlusion of the LAD. Thirty minutes later, the MSCs group received autologous MSCs transplantation through in?tramyocardial injection into the peri?infarcted areas (2×107,2 ml) and the control group was subjected to cell culture medium in the same way. At the 1st and 4th weeks after MSCs transplantation, myocardial glu?cose metabolism, myocardial perfusion and cardiac function were evaluated in the two groups through PET/CT, SPECT and MRI. The minimum FDG mean signal intensity ( MSI ) , summed MSI, SRS, SRS%, LVEF, ESV, stroke volume ( SV) and cardiac output ( CO) were calculated. On the 4th week, HE and Masson′s Trichrome stains were performed. Mann?Whitney u test and non?parametric Wilcoxon test were used. Results (1) As evaluated by PET in the 1st week, the MSI and summed MSI in MSCs group were less than those in control group ( 22. 10 ± 3. 18 vs 35. 70 ± 3. 02, z=-2. 65; 1 013. 50 ± 29. 37 vs 1 084. 00 ± 21?15, z=-1.97;both P0.05). (2) In the 1st week, the perfusion variables had no signifi?cant differences between the two groups ( P>0.05) . There was no significant difference in any perfusion vari?ables between the 1st and 4th weeks in the two groups, respectively (P>0.05). (3) As evaluated by MRI, the cardiac functional parameters had no significant differences between the two groups at the 1st week. In the MSCs groups, LVEF increased significantly ((54.41±2.62)% vs (47.54±2.43)%;z=-2.60, P0.05) . Conclusions Four weeks after MSCs transplantation for AMI, cardiac func?tion and myocardial glucose metabolism improved significantly but without significant myocardial perfusion improvement. Therefore, the cardiac function improvement might be associated with increased myocardial glucose metabolism.
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BACKGROUND:As a lipid-lowering drug, simvastatin has been proved to be effective in promoting bone formation. Previous studies have demonstrated that local y applied simvastation accelerated fracture healing at middle phase in osteoporotic rats, while no study focuses on the influence of local y applied simvastatin on fracture healing at late period in an osteoporotic rat. OBJECTIVE:To investigate the effect of simvastatin local y applied from a bioactive polymer coating of implants on osteoporotic fracture healing at late period. METHODS:Female Sprague-Dawley rats were divided into sham group, osteoporotic fracture group and simvastatin group. In the sham group, the abdominal cavity was exposed without ovariectomy. Six weeks later, femur fracture models were established in normal or osteotoporotic Sprague-Dawley rats, and intramedul ary stabilization was achieved with uncoated titanium Kirschner wires in normal rats (sham group),with polylactic acid coated titanium Kirschner wires (osteoporotic fracture group) and with simvastatin/polylactic acid coated titanium Kirschner wires (simvastatin group). Femurs were harvested after 12 weeks, bone mineral density was determined with dual X-ray absorptiometry, and then radiographic and histological analysis was performed for analysis of fracture healing. Immunohistochemical evaluation was employed for bone morphogenetic protein 2 expression. RESULTS AND CONCLUSION:The bone mineral densities of both the total fractured femur and fractured site 12 weeks after fracture in the osteoporotic fracture group and simvastatin group were markedly decreased compared to normal fractured rats. The bone mineral density of the fractured site was significantly higher in the simvastatin group than the osteoporotic fracture group. Radiographic results demonstrated completely finished cal us remodeling in the sham group;poor healing, pale cal us density and blurred fracture line were seen in the osteoporotic fracture group;disappearance of fracture line, bone defects fil ed with cal us, and deep periosteal reaction were found in the simvastatin group. X-ray scores in the sham and simvastatin groups were significantly higher than that in the osteoporotic group (P<0.05). Hematoxylin-eosin staining showed a delayed healing process in the osteoporotic group, and revealed a significantly processed cal us with regular-shaped newly formed bone trabeculae in the simvastatin group. Immunohistochemical evaluation showed no significant difference in the bone morphogenetic protein 2 expression between any two groups. These findings suggest an improved fracture healing under local application of simvastatin in osteoporotic rats.
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Objective To investigate the relationship between subclinical cervical HPV infection and recurrence of condyloma acuminatum (CA) in patients. Methods Cervical swabs were collected from 52 patients with frequently recurrent CA, 55 patients with infrequently recurrent CA, and 65 normal human controls. The Cenechip method was performed to detect the presence and type of HPV in cervical swabs followed by a statistical analysis. Results HPV was found in 67.3% (35/52) of swabs from patients with frequently recurrent CA, 5% (19/55) from patients with infrequently recurrent CA, and 4.6% (3/65) in the controls. There was a statistical difference among the three groups in the detection rate of HPV. Conclusion The subclinical cervical HPV infection may contribute to the recurrence of CA.
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AIM: To assess the degree of oxidative damage during acute myocardial infarction and reperfusion, and to clarify the protective effect of Tongxinluo in mini-swine model. METHODS: Thirty mini-swines were randomized into 5 study groups: sham group, model group, low dose (0.05 g·kg~(-1)·d~(-1)), medium dose (0.2 g·kg~(-1)·d~(-1)) and high dose (0.5 g·kg~(-1)·d~(-1)) of Tongxinluo groups (pretreated with Tongxinluo for 3 d). Animals except in sham group were subjected to 3 h of coronary occlusion followed by 1 h of reperfusion. Concentrations of total antioxidative capability (T-AOC), total superoxide dismutase (T-SOD), reduced glutathione (GSH) and malondialdehyde (MDA) in blood sample and the myocardium were measured. RESULTS: (1) T-AOC, T-SOD and GSH in serum significantly decreased (all P<0.05), while MDA significantly increased (P<0.01) at 3 h after AMI in comparison with those at baseline. Compared to those at 3 h after AMI, the contents of T-AOC, T-SOD and GSH at 1 h after reperfusion significantly decreased (all P<0.01), accompanied by increase of MDA (P<0.01). (2) Compared to those in normal area, levels of T-AOC, T-SOD and GSH in reperfusion myocardium decreased significantly (all P<0.01) and MDA increased significantly (P<0.01). T-AOC, T-SOD and GSH in no-reflow myocardium further decreased (all P<0.01) and MDA increased (P<0.01) as compared to those in reperfusion myocardium. (3) Compared to model group, medium dose of Tongxinluo increased the contents of T-AOC and T-SOD and reduced MDA production in serum at 3 h after AMI (all P<0.05), while medium dose of Tongxinluo increased T-SOD level at 1 h after reperfusion (P<0.05). High dose of Tongxinluo increased the levels of T-AOC and T-SOD and decreased MDA content in serum at 3 h after AMI and 1 h after reperfusion (all P<0.05). (4) The medium dose of Tongxinluo increased T-AOC content (P<0.05) and reduced MDA (P<0.05) in reperfusion myocardium, while high dose of Tongxinluo increased T-AOC, T-SOD and GSH (all P<0.05), reduced MDA (P<0.01) in reperfusion myocardium, and also increased T-AOC, T-SOD (all P<0.05), reduced MDA (P<0.01) in no-reflow area as compared to those in model group. CONCLUSION: Impairment of antioxidant defense system in vivo and imbalance of redox homeostasis in myocardium region might play an important role in the pathogenesis of no-reflow after myocardial acute infarction following reperfusion. Tongxinluo protects myocardium from reperfusion injury by improving antioxidant defense and attenuating oxidative damage.
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Objective To determine the concentrations of amphotericin B in serum and cerebrospinal fluid from patients with AIDS-associated cryptococcal meningitis.Methods Ten patients with AIDS-associated cryptococcal meningitis were enrolled in the study.Blood samples were collected before drug administrated at the 7th.14th and 21 th day after maintenance dose achieved.Cerebrospinal fluid samples were collected from lumbar puncture or taken before intrathecal drug administration at least one week after maintenance dose achieved.The concentrations of amphoteriein B in blood and cerebrospinal fluid samples were determined using RP-HPLC.Results When maintenance doses varied from 25 to 40 mg/d,the blood steady-state trough concentration range of amphotericin B was 1.34-2.27 ms/L,and that of cerebrospinal fluid was 44.0-9 1.7μg/L.Conclusions The concentrations of amphotericin B in the cerebrospinal fluid would be lower than the desired concentration if amphotericin B administrated only through intravenous or integrated with irregular intrathecal administration.It suggests that higher dosage of intravenous injection and regular intrathecal administration should be applied to achieve the desired concentration.
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Objective To investigate the ability of magnetic resonance imaging (MRI) in tracking magnetically labeled mesenchymal stem cells (MR-MSCs) in a swine myocardial infarction (MI) model.Methods Adult Chinese mini-pigs (n = 6) were subjected to open-chest experimental MI operation.Their antegeneic bone marrow-derived mesenchymal stem cells ( MSCs ) was cultured and doubly labeled with ferumoxides and DAPL On the 14 th day after MSCs transplantation, the size and location of the myocardial infarction were assessed by using delayed-enhancement MRI (DE-MRI). Then the labeled MSCs were injected intramyocardially into peri-infarct zone and normal myocardium. At 24 hrs and 3 weeks after injection, the contrast and the volume of the MR-MSCs hypointense lesion from the MR images were acquired, and the contrast was determined using the difference in signal intensity between the hypointense and normal myocardium divided by signal intensity of the normal region.After humane euthanasia, the heart was excised and histology corresponding to MRI slices that demonstrated MR-MSCs lesions was performed.Repeated-measures ANOVA and a paired t test were used for comparison of the contrast and the volume of the MR-MSCs hypointense lesion at different time points. Comparisons between independent groups were performed with the standard Student t test.Results The labeling efficiency of ferumoxides and DAPI was 100% . On the 14 th day after the MI operation, the average percentage of infracted myocardial area was (33.6±8.9)% .Twenty- four hours after MSCs transplantation, MSCs injection sites appeared as ovoid hypointensive lesions with sharp border on T2 * images. At 24 h after injection, the signal contrast [(67.00±5.48)% vs (61.92 ±7.76)%,t = 1.65,P =0.1158] and the size [(0.56 ±0.24) cm2 vs (0.52 ± 0.25 ) cm2, t = 0.39, P = 0.7044 ] of the lesions showed no statistical difference between the peri infarct zone and the normal myocardium.At 3 weeks after injection, the signal contrast decreased and the size diminished both in the peri-infarct zone and in the normal myocardium. Moreover, the contrast of the lesions in peri-infarct zone decreased more significantly than that in normal myocardium [(26.88 +7.27)%vs (15.00 :t:4.51)%, F =20.08, P =0.0003].Post mortem analysis found the fluorescontly labeled MSCs demonstrated on histological sections.There were much more dense fluorescently labled MSCs per high power fields at injection sites of normal myocardium than at injection sites of peri-infarct zone [ (106 ±25 )/HPF vs ( 143 ± 31 )/HPF, t = - 2.47, P = 0.0293 ].In MSCs injection sites of the peri-infarct zone,the capillary density was significantly higher than that in control sites [ (13.4 ± 4.0 )/HPF vs (9.4 ±3.1 )/HPF, t = 2.49, P = 0.0229].At 3 weeks after injection, ferumoxide was contained within partial original MSCs.Conclusion Magnetic resonance imaging of MSCs is a feasible method for the in vivo tracking of transplanted stem cells and could reflect the tendency of the local stem cell quantity, but there still has limitation for the semi-quantitation of the transplanted stem cells.
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During the reform of scientific research system the financial and the material resources were centralized and the central laboratory was constructed in cardiovascular Institute & Fu Wai Hospital.The biosafety was implemented thoroughly in this construction process.The brand-new concept war established and the strict rules were formulated through comprehensively study and repeatedly practice.In the meantime,the advanced management and the operation mode were established.A new biosafety lab was constructed under these conditions.
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OBJECTIVE:To determine the contents of chicoric acid in three different pharmaceutical dosage forms by RP-HPLC.METHODS:Samples were determined on VP-ODS C18,with the mobile phase consisted of methanol-1% acetic acid-tetrahydrofuran(33∶62∶5) with flow rate at 1.0 mL?min-1,UV detection wavelength at 327 nm,column temperature at 30 ℃ and sample size at 20 ?L.RESULTS:The linear range of chicoric acid was 0.398 72～3.987 2 ?g(r=0.999 8).The average recoveries of the oral liquid,capsules,and tablets were 99.85%(RSD=0.98%),102.50%(RSD=1.84%),and 100.50%(RSD=1.69%),respectively.CONCLUSION:This method is accurate,reproducible,specific,and suitable for the content determination of chicoric acid.
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To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.
Subject(s)
Cell Line, Transformed , Cloning, Molecular , Embryonic Structures , Eukaryotic Cells/metabolism , Gene Expression , Genetic Vectors , Kidney/cytology , Kidney/metabolism , Peroxidases/biosynthesis , Peroxidases/genetics , Plasmids/genetics , TransfectionABSTRACT
To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.
Subject(s)
Humans , Cell Line, Transformed , Cloning, Molecular , Embryo, Mammalian , Eukaryotic Cells , Metabolism , Gene Expression , Genetic Vectors , Kidney , Cell Biology , Metabolism , Peroxidases , Genetics , Peroxiredoxin III , Peroxiredoxins , Plasmids , Genetics , TransfectionABSTRACT
Objective:To investigate the correlation of dentofacial structure and the sagittal size of the upper airway in youthful nonsnoring men with malocclusion.Methods:Lateral cephalometry was carried out in 52 nonsnoring males with malocclusion. The average age of the patients was 19.23 years, with a range of 18 to 24 years. Stepwise regression analysis was used to relate those cephalometric parameters to the sagittal airway size at different levels.Results:SNB angle,S-N length and H-C_3 length were the related variables to superior posterior airway space(SPAS). H-C_3 length,SNB angle and L1-MP angle were the related variables to middle airway space(MAS). SNB angle,H-C_3 length,S-N length and L1-MP angle were the related variables to inferior airway space(IAS).Conclusion:The sagittal upper airway size in youthful nonsnoring men with malocclusion is correlated to the dentofacial structure and the hyoid position.