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Objective:To analyze the plague monitoring results of Marmota sibirica plague natural focus in Hulunbuir Plateau, and to study the characteristics of development and variation of the plague focus. Methods:From 2009 to 2020, monitoring was conducted in Manzhouli City, Prairie Chenbarhu Banner, Xin Barag Left Banner, and Xin Barag Right Banner from May to September each year. The one-day bow-clip method was used to investigate the density of the Spermophilus dauricus, and the 5-meter clamp method was used to investigate other small rodents. The classification and identification of ectoparasite fleas were done by using low power microscope. According to "Diagnostic Criteria for Plague" (WS 279-2008), the detectable materials of rats and fleas were tested, and Yersinia pestis was isolated and cultured. The serums of collected rats were detected by indirect hemagglutination test. Results:From 2009 to 2020, the average density of dauricus in Marmota sibirica plague natural focus of Hulunbuir Plateau was 0.45/hm 2, and the capture rate of other small rodents was 1.25% (787/63 091). A total of 9 species, 8 genera and 4 families of rodents were captured. The total flea infection rate of Spermophilus dauricus was 28.37% (1 945/6 856) and the fleas index was 1.04. A total of 3 species, 3 genera and 2 families of fleas were collected. A total of 6 674 Spermophilus dauricus were dissected and 6 787 fleas were cultured in 2 537 groups. The Yersinia pestis test was negative. A total of 6 795 serum samples from Spermophilus dauricus were detected, of which 11 were positive with a positive detection rate of 0.16%. The highest titer of positive serum was 1∶1 280. Indirect hemagglutination test positive materials were found for 5 consecutive years from 2010 to 2014; the positive serums were found in Manzhouli City and Xin Barag Right Banner. Conclusions:The Yersinia pestis antibady positive serum of Spermophilus dauricus has been detected in Marmota sibirica plague focus of Hulunbuir Plateau for many years, which is worthy of high vigilance. In the future, continuous surveillance should be carried out, bacterial detection should be strengthened, possible plague outbreaks should be detected in time, and various plague prevention and control should be done well.
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Objective To explore the effects of graphene oxide (GO)-polyethylene glycol (PEG)-folic acid (FA)-pyrenemethylamine hydrochloride (PyNH2)-mediated RNA interference (RNAi) of hypoxia-inducible factor-1α (HIF-1α) on the biological behaviors of human pancreatic cancer Patu8988 cells.Methods GO-PEG-FA-PyNH2 and RNAi targeting HIF-1α gene (GO-PEG-FA-PyNH2-HIF-1α-RNAi)was constructed.The expressions of HIF-1α and glucose transporter 1 (Glut-l) in Patu8988 cells were determined after knockdown of HIF-1α by RNAi.The invasive ability,the proliferation and the cell cycle of Patu8988 cells were investigated.The effect of HIF-1α knockdown on the uptake of 18F-fluorodeoxyglucose (FDG) in Patu8988 cells was also detected.Comparison of data was conducted by one-way analysis of variance and least significant difference t test.Results The GO-PEG-FA-PyNH2 was successfully constructed,and no cytotoxicity was found.Under the hypoxia or normoxia state,the mRNA and protein levels of HIF-1α and mRNA level of Glut-1 in cells transfected with GO-PEG-FA-PyNH2-HIF-1α-RNAi (study group) were lower than those in cells transfected with GO-PEG-FA-PyNH2 (negative group) and cells transfected with Opti-minimal essential medium (Opti-MEM,control group;F=30.25-32.58,t=3.66-5.81,all P＜0.05);the numbers of migrated cells in the study group were much lower than those in the negative group and the control group (F=38.63 and 41.35,t=20.51-35.25,all P＜0.01);the cell proliferation in the study group was significantly lower than that in the negative group and the control group (F=35.19 and 38.11,t =15.11-22.19,all P＜0.05).The proportions of G0/G1 cells in the study group were higher than those in the negative group and the control group (F=34.60 and 34.83,t=11.55-34.56,all P＜0.05);the 18 F-FDG uptake in the study group was lower than that in the negative group and control group (F=29.85 and 31.69,t =3.35-4.35,all P＜0.05).Conclusion GO-PEG-FA-PyNH2-mediated HIF-1α RNAi inhibits the expression of HIF-1α in pancreatic cancer cells,leading to changes in related biological behaviors.
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Objective To evaluate the predictive value of 18F-fluorodeoxyglucose (FDG) PET/CT before or after autologous hematopoietic stem cell transplantation (Auto-HSCT) in patients with non-Hodgkin's lymphoma (NHL).Methods Fifty-eight NHL patients who underwent PET/CT scan before or after Auto-HSCT were retrospectively analyzed.Between June 2007 and May 2016,40 patients (32 males,8 females;median age 39 years) underwent 18F-FDG PET/CT pre-transplantation,and 34 patients (23 males,11 females;median age 34 years) underwent 18F-FDG PET/CT post-transplantation.There were 16 patients underwent both pre-and post-transplantation imaging.PET/CT results were categorized as positive or negative according to the standard response criteria.The predictive value of PET/CT was evaluated with progression-free survival (PFS) and overall survival (OS) using Kaplan-Meier survival analysis and logrank test.Three-year PFS and OS of different groups were compared with Fisher exact test.Results (1) 18 F-FDG PET/CT before Auto-HSCT:PFS of patients in PET/CT negative group (n =27) and positive group (n=13) were significantly different (x2=4.187,P＜0.05),3-year PFS was 63.6％(14/22) vs 1/7 (P=0.031).However,OS of the 2 groups were not significantly different (x2=1.777,P＞0.05).(2)18F-FDG PET/CT after Auto-HSCT:PFS and OS of patients in PET/CT negative group (n=18) and positive group (n =16) were significantly different (x2 values:15.430,7.726,both P＜0.01).The 3-year PFS and OS of the 2 groups were 8/10 vs 2/12(P=0.005) and 8/10 vs 4/12 (P=0.038),respectively.Conclusion 18F-FDG PET/CT before or after Auto-HSCT could provide prognostic information for NHL patients.
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Objective To observe the antioxidant effects of 99Tc-MDP on the brain of aged mice induced by D-galactose.Methods A total of 48 healthy female Kunming mice were divided into 6 groups by random number table method:normal control group,model group,vitamin E (Vit E) group,groups treated with low,middle and high dosages of 99Tc-MDP.Except the normal control group,mice of each group were injected with 10％ D-galactose saline subcutaneously in the neck for 42 d.At the same time,the 99Tc-MDP groups were given different dosages of 99Tc-MDP (1.75× 10-5,3.50× 10-5,0.70× 10-4 μg/g according to the body mass) respectively into abdominal cavity twice a day.Vit E group was given Vit E by intragastric administration from the first day.The model group was injected with saline every day.After behavioral testing,mice serum samples and brain tissue samples were collected for the determination of superoxide dismutase (SOD),malondialdehyde (MDA),glutathione peroxidase (GSH-Px) and monoamine oxidase (MAO)levels.Then the mice were sacrificed and the brain tissue was taken for HE staining.The independent-sample t test was used for data analysis.Results The mice model was established successfully.SOD levels in brain tissue of groups injected with low,middle,high dosages of 99Tc-MDP,Vit E group were (19.06± 6.44),(20.41±4.02),(22.24±3.76),(24.71±5.09) U/mgprot,respectively,all of which were higher than that of the model group((11.32±2.90) U/mgprot;t=3.099 6-6.504 6,P＜0.05 or ＜0.01).There were similar results for GSH-Px (t =2.214 1-4.145 7,P＜0.05 or ＜0.01).MDA and MAO levels in brain tissue of 99Tc-MDP groups were lower than those of the model group (t =2.140 3-3.057 8,all P＜0.05).Compared to normal control group,the hippocampus in model group showed reduced cell number and layers,disordered structure,with part of the cells in smaller volume and abnormal nuclear shape.In Vit E group and the three 99Tc-MDP groups,no significant change of neuron was observed compared with normal control group,the degeneration and necrosis of hippocampal cells were mild compared with model group,the cell number and morphology were normal,and the structures were clear.Conclusion 99Tc-MDP may increase the activities of SOD,GSH-Px and reduce the levels of MDA and MAO in brain tissue of aged mice,thus it may be helpful in delaying the brain aging.
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Objective To evaluate the relationship between 18F-FDG uptake and P-gp expression in Bcap37 or Bcap37/MDR1 tumor-bearing BALB/c nude mice.Methods Bcap37 or Bcap37/MDR1 cells were injected into BALB/c nude mice (1× 107cells/ml,0.2 ml/mouse) to construct mice models.Bcap37 (n=5) or Bcap37/MDR1 (n=5) tumor-bearing mice fasted for 6 h before imaging.After anesthesia,the mice were injected with 7.4 MBq of 18F-FDG via tail vein.The dynamic microPET scans were carried out for 90 min.On the microPET images,the ROI was drawn and the TAC was obtained.The next day,those 10 mice underwent dynamic microPET scans after injected with elacridar (GF120918) and 18F-FDG.Another 10 mice,5 with Bcap37 tumors and 5 with Bcap37/MDR1 tumors,were used.After 7.4 MBq 18F-FDG with or without 2.0 mg/kg GF120918 was administered via tail vein,microPET images were acquired at 60 min.ROI was drawn over the tumors and SUV was obtained.Two-sample t test was used to analyze the data.Results GF120918 did not significantly alter the 18F-FDG accumulation curve in Bcap37 tumors,but significantly enhanced the 18F-FDG accumulation in Bcap37/MDR1 tumors.GF120918 did not influence 18F-FDG uptake (SUV) in Bcap37 tumors (1.052±0.028,1.028±0.045,t =1.792,P＞0.05),but significantly increased the SUV in Bcap37/MDR1 tumors (1.015±0.043,0.712±0.031,t=3.365,P＜0.05);The SUV of 18 F-FDG in Bcap37 tumors was significantly higher than that in Bcap37/MDR1 tumors without injection of GF120918 (t =3.952,P＜0.05).The SUV was not significantly different when GF120118 was injected (t=1.835,P＞0.05).Conclusions 18F-FDG is a substrate of P-gp.18F-FDG imaging combined with GF120918 injection may be an effective noninvasive method for the detection of tumor's MDR.
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Objective To evaluate the value of 18F-FDG PET/CT in histological grading and staging of follicular lymphoma (FL) and prediction of prognosis after first-line treatment.Methods From May 2007 to April 2013,24 patients (11 males,13 females; median age 55 years) with histopathological proof of FL were retrospectively studied.All patients were evaluated by 18F-FDG PET/CT before the first-line treatment and were divided into indolent FL and aggressive FL according to their histological grades and clinical stages.The sensitivity,specificity and SUVmax were calculated.Analysis of variance of factorial design was used to analyze the data.Within the same study period,18F-FDG PET/CT was also performed in 16 FL patients (9 males,7 females; median age 50.5 years) after first-line treatment (7/16 patients belonged to the aforementioned group with pretreatment PET/CT performed) ; and they were then divided into PET/CTpositive and PET/CT-negative groups.All 16 patients were followed for 6-49 months to evaluate the prognosis.The rates of overall survival (OS) and progression-free survival (PFS) were calculated.Results (1)The sensitivities of PET/CT in indolent and aggressive FL were 92.3％ (12/13) and 100％ (11/11),respectively.The SUVmax was 5.26± 1.70 vs 9.54±5.09 (F=5.196,P＜0.05).(2) According to PET/CT,3patients(12.5％,3/24) were upstaged from Ⅰ-Ⅱ to Ⅲ-Ⅳ,and 2 patients(8.3％,2/24) were downstaged from Ⅲ-Ⅳ to Ⅰ-Ⅱ.The SUVmax of stage Ⅰ-Ⅱ and Ⅲ-Ⅳ FL was 5.22±2.92 and 8.04±4.46(F=2.904,P＞0.05).(3)For the 16 FL patients with PET/CT after first-line treatment,the negative and positive groups had different OS and PFS.The 6-month OS,1-year OS and 3-year OS were 100％(13/13),9/9,4/5,respectively for the negative group,and 2/3,2/3,1/2,respectively for the positive group; while the corresponding 6-month PFS,1-year PFS and 3-year PFS were 92.3 ％ (12/13),8/9,3/5 and 2/3,0/3,0/2,respectively.Conclusions 18F-FDG PET/CT is valuable in the evaluation of histological grading and clinical staging of FL patients and in the prediction of prognosis after fist-line treatment.
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Objective To investigate the biodistribution and gamma imaging of 99Tcm-arginine-glutamate-threonine (RET) in nude mice bearing lung cancer xenografts and to explore its feasibility for human lung cancer imaging.Methods RET was labeled directly with 99Tcm.The binding efficiency of 99Tcm-RET with human NSCLC H1299 cells was measured.99Tcm-RET was injected via the tail vein in nude mice bearing H1299 xenografts.The mice(n=32) were sacrificed at different time points:15 min,30 min,1 h,2 h,4 h,8 h,24 h,and 48 h.Organs of interest were excised,weighted and counted by a gamma counter.The organ uptake was calculated as ％ID/g.The gamma imaging was performed on 3 mice at 0.5,1,2,4,4.5,5,6 h after intravenous injection of 4.81 MBq 99Tcm-RET.Results The radiolabeling yield of 99Tcm-RET was (93.15±2.02)％,and the binding efficiency of 99Tcm-RET with H1299 cells was (3.56±0.37)％.At 4 h after injection,the uptake of tumor,liver and spleen was (4.96±1.05) ％ID/g,(15.89±1.84) ％ID/g and (10.83±1.66) ％ID/g and the T/NT was 5.70±0.21 for the heart and 12.40±0.11 for the blood.The tumor in nude mice could be best visualized at 4.5-6.0 h.Conclusion 99Tcm-RET might have potential to serve as a lung cancer imaging agent.
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Objective To investigate the effect of 188Re-IGF-1 analogue (IGF-1A) in proliferation inhibition and apoptosis induction in human pancreatic carcinoma cell line Patu8988.Methods IGF-1A was labeled with 188Re.Patu8988 cells were divided into an un-treated control group,IGF-1A group (1,5,10,20 μg),188ReO4-group (0.37,1.85,3.70,7.40 MBq) and 188Re-IGF-1A group (0.37,0.74,1.85 MBq).The cell proliferation inhibition effects by the 188Re-IGF-1A and 188ReO4-were detected every day by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test from 1 d to 7 d after administration,while the IGF-1 A group was tested every day from 1 d to 6 d after treatment.Inhibition rates were calculated.At 3 d after treatment with 188ReO4-and 188Re-IGF-1A (1.85,3.70,7.40 MBq),cell apoptosis was detected by flow cytometry.For biodistribution studies of 188Re-IGF-1A,36 nude mice bearing Patu8988 cell xenografts were divided into6 groups.At different time points (15 min,1 h,4 h,1 d,3 d and5 d),36 mice (n =6 per time point) were sacrificed and organs of interest were removed,weighted and measured for radioactivity by a gamma counter.The absorbed doses of organs were calculated as ％ ID/g.One-way analysis of variance was used.Results After 4 d,inhibition rate of Patu8988 cell proliferation in the 188 Re-IGF-1A group (1.85 MBq) was (90.75 ±5.20) ％,higher than that in 188ReO4-group or IGF-1A group ((49.50±2.39)％,(23.00±4.21)％; F=554.724,P＜0.01).At 3 d after treatment with different doses of 188 Re-IGF-1A (1.85,3.70,7.40 MBq),floating cell ratios were (16.56 ± 0.95) ％,(33.39 ±5.93) ％ and (43.76 ± 1.38) ％,respectively.Apoptosisratios in the floating cells treated by 188 Re-IGF-1A (1.85,3.70,7.40 MBq) were (12.70±2.27)％,(17.80±1.51)％ and (23.23 ±1.22)％,respectively.Distribution in tumors was (39.30 ± 17.98),(10.59 ± 9.39),(5.32 ± 1.53) and (5.30 ±2.28) ％ ID/g at the 15 min,1 d,3 d,and 5 d timepoints after intratumoral injection,respectively.The absorbed dose of tumors was 5165.8 mGy/MBq.Conclusions Proliferation of human pancreatic carcinoma cell line Patu8988 can be inhibited and apoptosis can also be induced by 188Re-IGF-1A.The tumor region is the major distribution site in nude mice bearing human pancreatic cancer xenografts after intratumoral injection of 188 Re-IGF-1A.
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Objective To evaluate the effect of P-gp inhibitors of verapamil (VER) and GF120918 on 18F-FDG uptake in Bcap37 and Bcap37/multidrug resistancce (MDR)1 cell lines in vitro,and to explore the relationship between 18F-FDG uptake and P-gp expression at cellular level.Methods Bcap37 and Bcap37/MDR1 cells were seeded into 6-well plates at a density of 1 × 106 per well.Three days later,37 kBq/ml 18F-FDG,or 37 kBq/ml 18F-FDG + 100 μmoL/L VER,or 37 kBq/ml 18F-FDG + 50 μmol/L GF120918 were added into each well.Mter incubated for 10,30,60 and 120 min at 37 ℃ and in 5％ CO2,the medium was removed and the cells were washed three times with 1 ml ice-cold PBS immediately.The radioactivity of 18 F-FDG was measured using a gamma counter.The uptake of 18F-FDG was expressed as the ratio of 18F-FDG radioactivity in Bcap37 or Bcap37/MDR1 cells and the overall radioactivity added to the cells in each well.The t test was used for statistical analysis.Results 18F-FDG uptake was higher in Bcap37/MDR1 cells than that in Bcap37 cells after incubated for 10 min.The uptake rate was (1.88 ±0.19) ％ in Bcap37/MDR1 cells and (1.37 ± 0.18) ％ in Bcap37 cells (t =7.832,P ＜ 0.05).On the contrary,18 F-FDG uptake was significantly higher in Bcap37 cells than that in Bcap37/MDR1 cells after incubated for 60 and 120 min.The uptake rates were (2.29 ±0.23)％ and (2.34 ±0.15)％ in Bcap37 cells,(1.47 ±0.14)％ and (1.53 ±0.22)％ in Bcap37/MDR1 cells (t =8.437,8.283,both P ＜ 0.05).18 F-FDG uptake was significantly higher with VER or GF120918 in Bcap37/MDR1 cells than that without VER or GF120918 after the incubation of 60 and 120 min (t =9.032,9.243 and 8.765,8.803,all P ＜ 0.05).The uptake rates with VER or GF120918 were (2.45 ±0.21)％ and (2.46 ±0.25)％,(2.50 ±0.24)％ and (2.48 ±0.27)％.There was no significant difference of 18F-FDG uptake in Bcap37 cells with or without VER or GF120918.Conclusions 18F-FDG is a substrate of P-gp at cellular level.P-gp may act as an efflux pump to reduce 18F-FDG uptake in Bcap37/MDR1 cells.The uptake of 18F-FDG can be used to evaluate the function of P-gp in tumor cells.
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Objective To investigate the inhibitory effect of the lentiviral vector (LV)-mediated RNA interference (RNAi) targeting HIF-1α on the expression of HIF-1α and Glut-1 in human pancreatic cancer Patu8988 cells.Methods The RNAi targeting HIF-1α was combined to LV,and transfected into Patu8988 cells.The Patu8988 cells transfected with the empty vector and exposed to 0.5％ O2 for 4 h served as hypoxia negative control,the Patu8988 cells not transfected with vector and exposed to 0.5％ O2 for 4 h as hypoxia blank control,and the Patu8988 cells transfected with LV-RNAi-HIF-1α and exposed to 0.5％O2 for 4 h as experimental group.The expression of HIF-1α was measured by RT-PCR and Western blot respectively.The expression of Glut-1 was measured by RT-PCR.Each group was compared according to oneway analysis of variance and two-sample t test.Results After transfection with LV-RNAi-HIF-1α,HIF-1α mRNA expression decreased by 65.1％ (0.209/0.321) and 80.6％ (0.791/0.982) (t=10.52,15.24,both P＜0.05) under normoxia and hypoxia conditions,meanwhile with the empty vector,HIF-1α mRNA expression decreased by 0.6％ (0.002/0.321) and 7.2％ (0.071/0.982) (t =5.26,7.38,both P＜0.05).Under hypoxia conditions,the protein of HIF-1α in experimental group cells (0.159±0.010) was down-regulated obviously compared to the negative control group (0.745± 0.012) and the blank control group (0.711 ± 0.023)(F=35.52,t =6.72,10.56,all P＜0.05).The expression of Glut-1 mRNA in experimental group cells (0.040±0.003) decreased obviously compared to the negative control group (0.054±0.003) and blank control group (0.062±0.004) (F=35.28,t=5.94,8.55,all P＜0.01).Conclusion Gene silencing of HIF-1α using LV-mediated RNAi can inhibit the expression of HIF-1α and decrease the expression of Glut-1mRNA in Patu8988 cells.