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The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (Mpro) and papain like protease (PLpro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851Mpro inhibitors and 205 PLpro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both Mpro and PLpro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of Mpro inhibitors and over 20% of PLpro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 Mpro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Subject(s)
Humans , Antiviral Agents/chemistry , COVID-19 , COVID-19 Drug Treatment , High-Throughput Screening Assays , Molecular Docking Simulation , Protease Inhibitors/chemistry , SARS-CoV-2/enzymology , Viral Nonstructural ProteinsABSTRACT
Receptor activity-modulating proteins (RAMPs) are accessory molecules that form complexes with specific G protein-coupled receptors (GPCRs) and modulate their functions. It is established that RAMP interacts with the glucagon receptor family of GPCRs but the underlying mechanism is poorly understood. In this study, we used a bioluminescence resonance energy transfer (BRET) approach to comprehensively investigate such interactions. In conjunction with cAMP accumulation, Gα q activation and β-arrestin1/2 recruitment assays, we not only verified the GPCR-RAMP pairs previously reported, but also identified new patterns of GPCR-RAMP interaction. While RAMP1 was able to modify the three signaling events elicited by both glucagon receptor (GCGR) and glucagon-like peptide-1 receptor (GLP-1R), and RAMP2 mainly affected β-arrestin1/2 recruitment by GCGR, GLP-1R and glucagon-like peptide-2 receptor, RAMP3 showed a widespread negative impact on all the family members except for growth hormone-releasing hormone receptor covering the three pathways. Our results suggest that RAMP modulates both G protein dependent and independent signal transduction among the glucagon receptor family members in a receptor-specific manner. Mapping such interactions provides new insights into the role of RAMP in ligand recognition and receptor activation.
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We have previously shown that high expression of the nucleic acid binding factor YB-1 is strongly associated with poor prognosis in a variety of cancer types. The 3-dimensional protein structure of YB-1 has yet to be determined and its role in transcriptional regulation remains elusive. Drug targeting of transcription factors is often thought to be difficult and there are very few published high-throughput screening approaches. YB-1 predominantly binds to single-stranded nucleic acids, adding further difficulty to drug discovery. Therefore, we have developed two novel screening assays to detect compounds that interfere with the transcriptional activation properties of YB-1, both of which may be generalizable to screen for inhibitors of other nucleic acid binding molecules. The first approach is a cell-based luciferase reporter gene assay that measures the level of activation of a fragment of the promoter by YB-1. The second approach is a novel application of the AlphaScreen system, to detect interference of YB-1 interaction with a single-stranded DNA binding site. These complementary assays examine YB-1 binding to two discrete nucleic acid sequences using two different luminescent signal outputs and were employed sequentially to screen 7360 small molecule compounds leading to the identification of three putative YB-1 inhibitors.
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BACKGROUND:Nano-silver is a new antibacterial material developed based on nano-technology. It is characterized as stable physical and chemical properties, and good electrical, optical, and catalytic performance, but its antibacterial response to Staphylococcus aureus is controversial. OBJECTIVE:To explore the preparation methods of nano-silver coating on pure titanium surface and to explore its antibacterial effect on Staphylococcus aureus. METHODS:(1) Preparation of nano-silver coating on the surface of pure titanium: Hydroxyapatite and silver powder as research objects were fully mixed at a ratio of 20:1. The mixture was ball-milled to ensure the formation of nanosized particles. The mixture of hydroxyapatite and silver powder was put into a stirrer for continuous stirring to ensure the uniform distribution. The mixture was then placed in an ethanol solution followed by insertion of a titanium plate (anode) and a stainless steel plate (cathode). The precipitated powder was obtained on the titanium surface at a voltage of 20 V, and the pure titanium surface with nano-silver coating was prepared after heat treatment in a tube resistance furnace. (2) Observation of antibacterial properties: A minimal inhibitory concentration test was used to determine the antibacterial concentration of the nano-silver coating on the pure titanium surface when shaken with Staphylococcus aureus. A scanning electron microscope was used to observe the structure of Staphylococcus aureus on the pure titanium surface with nano-silver coatings of different mass concentrations. RESULTS AND CONCLUSION:Nano-silver/hydroxyapatite composite coating and hydroxyapatite coating were relatively uniform. The nano-silver/hydroxyapatite composite coating was white in color but slightly yellowish. The hydroxyapatite coating was white in color. The coating surface was rough and bonded firmly. No peeling of the coating was observed under gross observation. The antibacterial ability of the nano-silver group at 37℃ static culture and at 37℃ shaking culture was significantly higher than that of the hydroxy-apatite group (P<0.05). The absorbance value of Staphylococcus aureus at 600 nm in the nano-silver group was lower than that of hydroxyapatite group at 7, 10, 30 hours after intervention (P<0.05). The layer cells on the nano-silver coating become lighter in color compared with those on the hydroxyapatite coating and there were cells that ruptured and died. The number of Staphylococcus aureus on the nano-silver coating was reduced, and a large number of vacuoles were found. These findings indicate that the 20:1 mixture of hydroxyapatite and silver at micron level can be used to prepare nano-silver coating through ball milling, water bath, ultrasound and heat treatment. The prepared coating can exert excellent antibacterial effects on Staphylococcus aureus.
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Interferons (IFNs) are cytokines with fundamental roles in resistance to infections, cancer and other diseases. Type-I IFNs, interferon (IFN-) and interferon (IFN-), act through a shared receptor complex (IFNAR) comprised of IFNAR1 and IFNAR2 subunits. Binding of type-I IFN to IFNAR1 will robustly activate Janus activated kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway. Aberrant activation of the type-I IFN response results in a spectrum of disorders called interferonopathies. The purpose of this research is to develop an assay for high-throughput screening (HTS) of small molecule inhibitors of the type-I IFN signaling pathway. Inhibition of type-I IFN signaling can be beneficial in terms of therapeutic use and understanding the underlying mechanism of action. We report here a HTS campaign with the secreted embryonic alkaline phosphatase (SEAP) reporter gene assay against 32,000 compounds which yielded 25 confirmed hits. These compounds were subsequently characterized for their cytotoxicity, effects on STAT phosphorylation and activities in IFN regulatory factor (IRF) transcription.
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The serum and glucocorticoid inducible protein kinase (SGK) family members share similar structure, substrate specificity and function with AKT and signal downstream of the phosphatidylinositol 3-kinase (PI3K) signalling pathway. They regulate a range of fundamental cellular processes such as cell proliferation and survival, thereby playing an important role in cancer development. This perspective intends to give an overview on the involvement of SGKs (particularly SGK3) in cancer progression, and compares the actions of SGK3 and AKT in cell cycle regulation, oncogenic signalling, and the potential as a therapeutic target for cancer.
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Objective To observe the lung injury of rats caused by PM2.5 and the intervention effect of Platycodonis Radix total saponins (PGS); To discuss the repair mechanism of PGS for lung injury caused by PM2.5. Methods The model of lung injury caused by PM2.5 was induced by tracheal instillation. The rats were randomly divided into blank group, model group and PGS high-, medium- and low-dose groups, with 10 rats in each group. After medication for 14 d, BALF and left lung and right lung tissue of rats were collected. HE staining was used to observe the pathological changes of lung tissue. The contents of TNF-α, IL-6, IL-10 and IL-13 in BALF were detected by ELISA. RT-PCR was used to detect the expression of TGF-β mRNA in lung tissues. The expression of TGF-β protein in lung tissue was detected by Western blot. Results Compared with model group, TNF-α and IL-6 decreased significantly while IL-10 and IL-13 increased significantly in BALF in high- and medium-dose group of PGS. Degree of pulmonary interstitial edema and fibroplasia alleviated in PGS groups. Expressions of TGF-βmRNA and protein in lung tissue was reduced in PGS groups. Conclusion PGS can alleviate inflammatory injury caused by PM2.5 via regulating the balance of cytokine and down-regulating the expression of TGF-β and inhibiting the development of fibrosis, which results in the protection and repair on the lung injury of rats caused by PM2.5.
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Objective To understand the situation and epidemic characteristics of imported falciparum malaria in Huai'an City from 2010 to 2016,so as to provide the evidence for formulating the prevention and control strategies of imported falci-parum malaria in the city. Methods The epidemic data of imported falciparum malaria in Huai'an City from 2010 to 2016 were analyzed by using the descriptive epidemiological method. Results A total of 308 malaria cases were reported in Huai'an City from 2010 to 2016 with the average annual incidence of 0.88/105. A total of 240 imported falciparum malaria cases were report-ed,of which 18 cases(7.50%)developed into severe illness,and 2 severe patients died. The cases were reported in every coun-ty(district),and the incidence rates of Qingpu District and Huai'an District were higher than the city average level. The cases occurred every month,so there was no significant seasonal variation in the reporting time of the cases. Most of the patients were young men and aged 30-49 years. The occupational distribution revealed that the patients were mainly farmers,workers and mi-grant workers. The main source of infection was from African countries. The median interval from symptom appearing to definite diagnosis was 1 day,and the longest interval was 236 days. Twenty-nine cases were diagnosed within 24 hours,accounting for 12.08%. Conclusions The epidemic situation of imported falciparum malaria in Huai'an City is grim. In order to consolidate the achievements of malaria eradication,it is necessary to further improve the multi-sectoral cooperation mechanism,strengthen the management of floating population and take effective measures to reduce the risk of imported falciparum malaria.
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Objective·To screen NPHS2 mutations in adult focal segmental glomerulosclerosis(FSGS)patients based on a large Chinese FSGS cohort. Methods · All patients were biopsy determined FSGS by the Department of Nephrology at Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine. FSGS secondary to systemic disease and other hereditary kidney disease were excluded. After extraction of genomic DNA of peripheral blood,NPHS2 was screened by directly sequencing the exon/intron junction or high-throughput sequencing,including whole exon sequencing and Panel sequencing, and then verified by Sanger sequencing. One hundred healthy controls were enrolled to validate candidate mutations. Results · Two hundred and four FSGS patients were enrolled,including 52 familial(25.5%) and 152 sporadic patients(74.5%),of which steroid-resistant FSGS patients accounted for 30.3%(46/152).By sequencing NPHS2 in all patients of the cohort(Sanger sequencing in 61 patients and high-throughput sequencing in 143 patients), 2 novel conserved mutations were identified, one homozygous mutation in sporadic steroid-resistant FSGS, p.N199I and one heterozygous mutation in familial FSGS, p.L321fx346. Both of them were not detected in 100 healthy controls. These two variants were predicted to be damaging by Polyphen,SIFT and Mutation Taster.Totally,the mutation rate of NPHS2 in the FSGS cohort was 1%. Conclusion·Since the overall frequency of NPHS2 mutations is considerably low in Chinese adult-onset FSGS,NPHS2 is not the main disease-causing gene of this group of people.
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Objective@#To investigate the safety and effectiveness of radical retropubic prostatectomy (RRP) with adjuvant androgen deprivation or external radiotherapy in the treatment of prostate cancer (PCa) with pelvic lymph node metastasis (PLNM).@*METHODS@#Twenty PCa patients underwent bilateral pedal lymphangiography (PLG) preoperatively, and 11 of them received lymph node aspiration for examination of the mRNA expressions of prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSMA) in the lymph fluid by real-time RT-PCR. All the patients were treated by RRP with extended dissection of pelvic lymph nodes, and 3 of them by external radiotherapy in addition after recovery from urinary incontinence because of positive surgical margins, followed by adjuvant androgen deprivation therapy.@*RESULTS@#Real-time RT-PCR showed positive mRNA expressions of PSA and PSMA in the lymph fluid of the 11 patients, all pathologically confirmed with PLNM. The median intraoperative blood loss was 575 ml, with blood transfusion for 5 cases. Positive surgical margin was found in 3 cases, lymphorrhagia in 2 and urinary leakage in another 2 each. There were no such severe complications as vascular injury and rectum perforation. The patients were followed up for 6－48 (mean 42) months, during which, biochemical recurrence was observed in 12 cases at a median of 12 months postoperatively and 2 patients died at 12 and 48 months respectively.@*CONCLUSIONS@#Bilateral PLG and lymph node aspiration for examination of the mRNA expressions of PSA and PSMA in the lymph fluid help to confirm PLNM preoperatively. Radical retropubic prostatectomy with adjuvant androgen deprivation or external radiotherapy is safe and effective for the treatment of PCa with PLNM, but it should be chosen cautiously for those with Gleason 5+5.
Subject(s)
Humans , Male , Androgen Antagonists , Therapeutic Uses , Antigens, Surface , Metabolism , Chemotherapy, Adjuvant , Glutamate Carboxypeptidase II , Metabolism , Lymph Node Excision , Lymph Nodes , Pathology , Lymphatic Metastasis , Pelvis , Postoperative Period , Prostate-Specific Antigen , Metabolism , Prostatectomy , Methods , Prostatic Neoplasms , Drug Therapy , Metabolism , General SurgeryABSTRACT
Eukaryotic elongation factor 2 kinase (eEF2K) inhibitors may aid in the development of new therapeutic agents to combat cancer. Purified human eEF2K was obtained from anexpression system and a luminescence-based high-throughput screening (HTS) assay was developed using MH-1 peptide as the substrate. The luminescent readouts correlated with the amount of adenosine triphosphate remaining in the kinase reaction. This method was applied to a large-scale screening campaign against a diverse compound library and subsequent confirmation studies. Nine initial hits showing inhibitory activities on eEF2K were identified from 56,000 synthetic compounds during the HTS campaign, of which, five were chosen to test their effects in cancer cell lines.
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Systemic lupus erythematosus (SLE) and clear cell renal cell carcinoma (CC-RCC) are serious disorders and usually fatal, and always accompanied with pathological changes in the kidney. Signal-induced proliferation-associated protein 1 (SIPA-1) is a Rap1GTPase activating protein (Rap1GAP) expressed in the normal distal and collecting tubules of the murine kidney. Lupus-like autoimmune disease and leukemia have been observed in SIPA-1 deficient mice, suggesting a pathological relevance of SIPA-1 to SLE and carcinoma in human being. The expression pattern of SIPA-1 is as yet undefined and the pathogenesis of these diseases in humans remains elusive. In this study, we used both immunohistochemistry and quantum dot (QD)-based immunofluorescence staining to investigate the expression of SIPA-1 in renal specimens from SLE and CC-RCC patients. MTT assay and Western blotting were employed to evaluate the effects of SIPA-1 overexpression on the proliferation and apoptosis of renal cell lines. Semi-quantitative reverse transcriptase-PCR (RT-PCR) was applied to examine the changes of hypoxia-inducible factor-1α (HIF-1α) mRNA level. Results showed that SIPA-1 was highly expressed in the proximal and collecting tubules of nephrons in SLE patients compared to normal ones, and similar results were obtained in the specimens of CC-RCC patients. Although SIPA-1 overexpression did not affect cellular proliferation and apoptosis of both human 786-O renal cell carcinoma cells and rat NRK-52E renal epithelial cell lines, RT-PCR results showed that HIF-1α mRNA level was down-regulated by SIPA-1 overexpression in 786-O cells. These findings suggest that SIPA-1 may play critical roles in the pathological changes in kidney, and might provide a new biomarker to aid in the diagnosis of SLE and CC-RCC.
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Systemic lupus erythematosus (SLE) and clear cell renal cell carcinoma (CC-RCC) are serious disorders and usually fatal, and always accompanied with pathological changes in the kidney. Signal-induced proliferation-associated protein 1 (SIPA-1) is a Rap1GTPase activating protein (Rap1GAP) expressed in the normal distal and collecting tubules of the murine kidney. Lupus-like autoimmune disease and leukemia have been observed in SIPA-1 deficient mice, suggesting a pathological relevance of SIPA-1 to SLE and carcinoma in human being. The expression pattern of SIPA-1 is as yet undefined and the pathogenesis of these diseases in humans remains elusive. In this study, we used both immunohistochemistry and quantum dot (QD)-based immunofluorescence staining to investigate the expression of SIPA-1 in renal specimens from SLE and CC-RCC patients. MTT assay and Western blotting were employed to evaluate the effects of SIPA-1 overexpression on the proliferation and apoptosis of renal cell lines. Semi-quantitative reverse transcriptase-PCR (RT-PCR) was applied to examine the changes of hypoxia-inducible factor-1α (HIF-1α) mRNA level. Results showed that SIPA-1 was highly expressed in the proximal and collecting tubules of nephrons in SLE patients compared to normal ones, and similar results were obtained in the specimens of CC-RCC patients. Although SIPA-1 overexpression did not affect cellular proliferation and apoptosis of both human 786-O renal cell carcinoma cells and rat NRK-52E renal epithelial cell lines, RT-PCR results showed that HIF-1α mRNA level was down-regulated by SIPA-1 overexpression in 786-O cells. These findings suggest that SIPA-1 may play critical roles in the pathological changes in kidney, and might provide a new biomarker to aid in the diagnosis of SLE and CC-RCC.
Subject(s)
Humans , Apoptosis , Base Sequence , Cell Line , Cell Proliferation , DNA Primers , GTPase-Activating Proteins , Metabolism , Kidney Tubules, Proximal , Metabolism , Pathology , Lupus Erythematosus, Systemic , Metabolism , Pathology , Nuclear Proteins , Metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To explore the clinical significance of cytokeratin 19 fragments test in the diagnosis of nasopharyngeal carcinoma.</p><p><b>METHODS</b>The study included 102 cases of nasopharyngeal carcinoma, 90 cases of nasal polyp/nasopharyngitis, and 150 healthy individuals. RT-PCR was used to detect CK19 mRNA expression and Western blot to detect CK19 fragment protein expression in tissues of nasopharyngeal carcinoma. Expression of CK19-2G2 was examined by immunohistochemistry. Chemiluminescence analysis was used to detect the serum levels of CK19-2G2, and ELISA to detect that of EB-VCA IgA.</p><p><b>RESULTS</b>Among 102 cases of nasophryngeal carcinoma, 64 showed CK19 mRNA expression by RT-PCR, 60 showed CK19 protein fragments in tumor tissues by Western blot, and 66 showed expression of CK19-2G2 by immunohistochemistry in nasopharyngeal carcinoma, including strong positivity in 20 cases, moderate in 34 cases and weak in 12 cases. The sensitivity and specificity of CK19-2G2 in the diagnosis of nasopharyngeal carcinoma were 49.0% and 89.2%, and those of EB-VCA IgA were 52.9% and 85.4%, respectively. The combined detection of CK19-2G2 and EB-VCA IgA increased the sensitivity to 73.5% while the specificity remained at 80.0%.</p><p><b>CONCLUSIONS</b>High levels of CK19-2G2 fragment expressed in tissue and serum are present in patients with nasopharyngeal carcinoma. The serum level of CK19-2G2 is helpful in the diagnosis of nasopharyngeal carcinoma. Furthermore, the combination of serum CK19-2G2 and EB-VCA IgA improves the detection sensitivity.</p>
Subject(s)
Adult , Aged , Humans , Middle Aged , Antigens, Viral , Blood , Blotting, Western , Capsid Proteins , Blood , Carcinoma, Squamous Cell , Diagnosis , Metabolism , Pathology , Herpesvirus 4, Human , Allergy and Immunology , Immunoglobulin A , Blood , Immunohistochemistry , Keratin-19 , Blood , Metabolism , Nasopharyngeal Neoplasms , Diagnosis , Metabolism , Pathology , Neoplasm Staging , Peptide Fragments , Blood , Metabolism , RNA, Messenger , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and SpecificityABSTRACT
<p><b>BACKGROUND</b>Improving the success rate of ureteroscopic lithotripsy for proximal ureteral stones is the hot issue in this field. Here we reported our experience on the treatment of proximal ureteral stones.</p><p><b>METHODS</b>From 2005 to 2010, 187 consecutive patients with proximal ureteral stones who underwent ureteroscopic lithotripsy were enrolled. The initial 52 patients treated by semi-rigid ureteroscope alone were classified as group 1. The subsequent 135 patients treated by semi-rigid ureteroscope with the aid of stone basket and flexible ureteroscope were classified as group 2.</p><p><b>RESULTS</b>In group 1, the overall stone-free rate was 67.3%. By a single procedure of ureteroscopic lithotripsy using a semi-rigid instrument, patients with ureteral stones below the 4th lumbar vertebra level achieved 91.7% stone-free rate, which was only 50% in patients with stones above the 4th lumbar vertebra level. Conversion to open surgery occurred in two patients since ureteral perforation was observed. In group 2, the stone-free rate achieved 93.2% with the aid of an N-Trap basket, which was significantly higher than that of patients without the aid of the basket (51.6%). Flexible ureteroscope was subsequently used in patients with fragment migration, thus making the overall success rate in group 2 increases to 97.0%.</p><p><b>CONCLUSIONS</b>Ureteroscopic lithotripsy is a safe and efficacious treatment for proximal ureteral stones. A single procedure of ureteroscopic lithotripsy using semi-rigid ureteroscope could achieve a satisfactory stone-free rate in patients with proximal ureteral stones below the 4th lumbar vertebra level. However, patients with ureteral stones above the 4th lumbar vertebra level experienced higher stone-migration rate, which would decrease the success rate. Fortunately, the stone-free state could possibly be achieved with the aid of an N-trap basket and flexible ureteroscope.</p>
Subject(s)
Adolescent , Adult , Aged , Humans , Middle Aged , Young Adult , Lasers, Solid-State , Therapeutic Uses , Lithotripsy, Laser , Methods , Ureteral Calculi , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To search for an effective method of reducing intraoperative blood loss in radical retropubic prostatectomy (RRP).</p><p><b>METHODS</b>We performed RRP for 100 patients with prostate cancer, 50 (group A) with the Walsh or Poor method for handling the dorsal venous complex (DVC), and the other 50 (group B) through the following three additional procedures for hemostasis: first placing a #7 prophylactic suture in the distal position of DVC, then ligating the vascular bundle of the prostatic apex with continuous 4-0 Vicryl sutures, and lastly placing a 4-0 absorbable suture followed by freeing the neurovascular bundle (NVB) or freeing NVB before suturing the remained levator ani myofascia and the deep layer of Denovilliers' fascia above the rectal serosa with 4-0 Vicryl. We assessed the effects of the three hemostatic methods in RRP by comparing the volumes of intraoperative blood loss and transfusion, operation time and perioperative levels of hemoglobin.</p><p><b>RESULTS</b>There were no significant differences between groups A and B in age, PSA, Gleason score, clinical stage, prostate volume, operation time and perioperative hemoglobin levels (P>0.05). The volumes of intraoperative blood loss and transfusion were markedly higher in group A ([1103.00 +/- 528.03] ml and [482.00 +/- 364.60] ml) than in B ([528.00 +/- 258.96] ml and [140.00 +/- 266.28] ml) (P<0.05).</p><p><b>CONCLUSION</b>Intraoperative blood loss in RRP could be significantly decreased by placing a prophylactic hemostatic suture in the distal position of DVC, continuous suture of the vascular bundle of the prostatic apex after cutting off the urethra, and placing a fine absorbable suture above NVB or continuous suture of the remained levator ani mony fascia and the deep layer of Denovilliers'fascia above the rectal serosa with absorbable sutures after freeing NVB.</p>
Subject(s)
Aged , Humans , Male , Middle Aged , Blood Loss, Surgical , Hemostatic Techniques , Prostatectomy , Methods , Prostatic Neoplasms , General SurgeryABSTRACT
Objective To evaluate 18F-N- succinimidyl -4-fluorobenzoate (SFB)-Annexin B1 in detectingin vitro andin vivo apoptosis. Methods Anti-Fas antibody was used to induce apoptosis in Jurkat cells. Apoptosis in Jurkat cells was confirmed by flow cytometer (FCM). Unilateral renal ischemia/reperfusion injury was induced by transient (45 min) ligation of the renal artery in the rabbit. The rabbit was then administrated with 18F-SFB-Annexin B1 intravenously 24 h later and then imaged by PET/CT at 10,30,60,90,120 and 240 min postinjection. Apoptosis in kidney was confirmed by terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) assay and HE staining. Results The apoptosis rate induced by anti-Fas antibody was 25.98%(120 min) while that in the control group was only 1.81%. The uptake of 18F-SFB-Annexin B1in apoptosis group was greater than that in the control group. PET/CT images at 240 min showed higher uptake in the ligated kidney than the non-ligated kidney. TUNEL assay and HE staining confirmed great amount apoptotic cells in the ligated kidney. Conclusion 18 F-SFB-Annexin B1may be potentially useful in detecting apoptosis both in vitro and in vivo.
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Objective To synthesize 16α-[18F]fluoro-17β-estrogen(18F-FES) with high chemical and radiochemical purity using multifunctional radiolabeling module of Explora GN and separation module of Explora LC (Explora GN/LC dual module). Methods Explora GN/LC dual module and one-pot radiochemical process were applied for fully automated synthesis and separation of 18F-FES. The radiochemical purity of 18F-FES was analyzed by HPLC. The saturation binding test of 18F-FES with ER (ER-positive MCF-7 cell line) was applied to examine the biological activity of 18F-FES. Results 18F-FES synthesis procedure took 60 min, with radiochemical yield about 45% and radiochemical purity higher than 98%. The Kd and Bmax of MCF-7 cells with 18F-FES were (2.922±0.619) nmol/L and (4.193±0.360) nmol/L respectively. Conclusions 18F-FES can be automatically synthesized by Explora GN/LC dual module with high chemical purity and radiochemical purity. It would be a potential PET tracer to be used in estrogen-related molecular imaging.
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<p><b>OBJECTIVE</b>To summarize the experience and lessons from 100 cases retropubic radical prostatectomy performed in the past 10 years.</p><p><b>METHODS</b>From July 1999 to July 2009, we performed 100 cases of retropubic radical prostatectomy, of which 84 were followed up for 3 - 120 months. We analyzed their preoperative age, PSA level, amount of intraoperative blood transfusion, operation time, urinary continence, penile erectile function, stricture of the anastomotic stoma and Qmax.</p><p><b>RESULTS</b>The mean age, PSA level, amount of intraoperative blood transfusion, operation time were 66.8 yr, 20.1 ng/ml, 585.7 ml and 198.9 min; the recovery rates of bladder control at 3, 6 and 12 months postoperatively were 65.5%, 81.7% and 92.4%, respectively. At 12 months after surgery, penile erection was restored in 19 cases (42.2%), anastomotic stoma stricture developed in 5 (5.9%), Qmax averaged 20.5 ml/min, biochemical recurrence was found in 13, and 1 died from prostate cancer.</p><p><b>CONCLUSION</b>Retropubic radical prostatectomy is a desirable procedure for the treatment of local prostate cancer, in which ligation of the puboprostatic ligament and prostatic venous plexus before cutting off the ligament helps improve urinary continence, protection of the neurovascular bundle and collateral pudendal artery contributes to the recovery of penile erectile function, and proper connection of urethral and bladder mucosa can reduce anastomotic stoma stricture. Postoperative external-beam radiotherapy for those with T3a or local lymph node metastasis could decrease biochemical recurrence.</p>
Subject(s)
Aged , Humans , Male , Middle Aged , Follow-Up Studies , Prostatectomy , Methods , Prostatic Neoplasms , General SurgeryABSTRACT
<p><b>OBJECTIVE</b>To clarify the significance of micrometastases in pelvic lymph nodes in patients with neoadjuvant hormonal therapy (NHT) before radical prostatectomy (RP).</p><p><b>METHODS</b>Twenty-one patients with clinically localized prostate cancer who received NHT between August 2007 and March 2010 were observed. The patients were clarified into four groups: pathological examination was positive (group A), real-time PCR examination targeting prostate specific antigen (PSA) mRNA and prostate specific membrane antigen (PSMA) mRNA were positive (group B), pathological examination and real-time PCR examination targeting PSA mRNA and PSMA mRNA were both negative (group C), and the control group (group D). After a standard bipedal lymphangiography the films were reviewed carefully by an experienced radiologist. If positive lymph nodes were seen or suspected, a thin-walled 22 gauge needle were directed transabdominally under fluoroscopic control into the area of question and an aspirate was obtained. The expression of PSA and PSMA in aspirate were assessed by a fully quantitative real-time PCR. The specimens were regarded in which either PSA mRNA or PSMA mRNA were positive as showing the "presence of micrometastasis". Lymph node specimens were also stained immunohistochemically with an antibody PSA after RP.</p><p><b>RESULTS</b>Pathological examination detected lymph node metastases from 3 cases, and real-time PCR further identified lymph node micrometastases from 14 cases with no pathological evidence of nodal involvement. The expression level of PSA mRNA and PSMA mRNA were statistically significant in patients with histological confirmed lymph node metastases and micrometastases detected by real-time PCR despite the lack of histological evidence, and the expression level of PSA mRNA and PSMA mRNA in aspirate were higher than the lymph node between the group A and group B.</p><p><b>CONCLUSIONS</b>Although residual foci of atrophic prostate cancer cells in resected lymph nodes after NHT can be difficult to diagnose by pathological examination, the present results show the usefulness of quantitative real-time PCR targeting PSA and PSMA mRNA for detected micrometastatic tumour foci in pelvic lymph nodes from fine needle aspiration biopsy of lymph nodes before RP.</p>