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Objective:To evaluate the effect of ladder teaching of problem-based learning (PBL) after theoretical lecture in the standardized training of ophthalmology.Methods:This study collected the data of the resident trainees in the ophthalmology standardized training base of the PLA General Hospital from September 2015 to June 2020. According to the accepted teaching methods, they were divided into the ladder teaching method group (ladder group) and the pure PBL teaching method group (PBL group). The exam results of the first and second stages of the trainees, the pass rate of the first stage exam (ophthalmology theory), the pass rate of the second stage exam (clinical application), and the overall pass rate of the first and second stages were recorded for statistical analysis. Quantitative evaluation on the teaching effect of the "ladder teaching method of theory teaching followed by PBL" in the standardized training of ophthalmology in this training base was made. SPSS 24.0 was used to conduct t-test, rank-sum test and Chi-square test. Results:The results of the first stage examination (ophthalmology theory) in the ladder group were higher than those in the PBL group [(87.22±8.45) vs. (74.47±10.68)], with a statistically significant difference ( P < 0.01); the pass rate of the first stage examination (ophthalmology theory) in the ladder group was higher than that in the PBL group (95.83% vs. 85.00%), with no significant difference ( P = 0.213). The pass rate of the second stage examination (clinical application) in the ladder group was higher than that in the PBL group (95.65% vs. 76.47%), with no significant difference ( P = 0.070); the overall pass rate of routine training (first and second stages) in the ladder group was higher than that in the PBL group (91.67% vs. 65.00%), with a statistically significant difference ( P = 0.029). Conclusion:The ladder teaching of PBL after the theory teaching has a satisfactory effect in the standardized training of ophthalmology. This teaching method is easier to help the students to successfully complete the standardized training and pass the standardized training examination.
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Aim To investigate the effects of long non-coding RNA(lncRNA)UNC5B-AS1 on the proliferation and epithelial mesenchymal transformation(EMT)of cervical cancer. Methods GEO and TCGA databases were used to download data sets and differential expression analysis was performed. qRT-PCR was used to verify the differential expression of lncRNA UNC5B-AS1 in normal and cancerous cervical tissues.The interference and overexpression of lncRNA UNC5B-AS1 were transfected into cervical cancer cell lines, and plate cloning, CCK-8 and EdU experiments were used to detect the effect of lncRNA UNC5B-AS1 on the pro-liferation of cervical cancer cells.Transwell assay was used to detect its effect on migration and invasion of cervical cancer cells.The expression levels of EMT-related genes E-Cadherin, N-Cadherin and Vimentin were detected by Western blot. Transcriptome sequencing was used to obtain the signal pathway regulated by lncRNA UNC5B-AS1, and to verify the expression level of related genes. Results RNA microarray and bioinformatics analysis showed that the expression level of lncRNA UNC5B-AS1 in cervical cancer was significantly higher than that in normal cervical tissue, and correlated with the overall survival time of patients.Compared with the negative control group, knockdown lncRNA UNC5B-AS1 could reduce the proliferation, migration and invasion of cervical cancer cells, while overexpression could promote the proliferation, migration and invasion of cervical cancer cells. Western blot showed that lncRNA UNC5B-AS1 could regulate EMT of cervical cancer cells. Transcriptome sequencing showed that lncRNA UNC5B-AS1 could regulate Toll like receptor(TLR)signaling pathway. qRT-PCR and Western blot results showed that the expression levels of TLR-related genes IL-6 and TICAM2 in the knockdown and overexpression lncRNA UNC5B-AS1 group were significantly changed(P<0.05). Conclusions LncRNA UNC5B-AS1 is highly expressed in cervical cancer. Overexpression of lncRNA UNC5B-AS1 may enhance TLR signaling pathway activity, thereby promoting proliferation and EMT of cervical cancer cells.
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Mother-to-child transmission of hepatitis B virus (HBV) is one of the main ways of transmission and the main cause of chronic hepatitis B after infection. Therefore, preventing mother-to-child transmission of HBV is particularly important in reducing the incidence of chronic hepatitis B. Currently, nucleoside ( acid) analoids ( Nas ) used for mother-to-child blocking of HBV include lamivudine (LAM) , tibivudine (LdT) and tenofovir fumarate ( TDF). Propofol tenofovir fumarate (TAF) has also been used in pregnant chronic hepatitis B pa- tients. This paper summarizes the efficacy, safety and antiviral treatment indications and termination time of the above-mentioned drugs in mother-to-child preventing to provide suggestions for the selection and rational application of mother-to-child preventing Nas.
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Parkinson's disease (PD) is the second most common and fastest-growing neurodegenerative disorder. In recent years, it has been recognized that neurotransmitters other than dopamine and neuronal systems outside the basal ganglia are also related to PD pathogenesis. However, little is known about whether and how the caudal zona incerta (ZIc) regulates parkinsonian motor symptoms. Here, we showed that specific glutamatergic but not GABAergic ZIc
Subject(s)
Animals , Mice , Neurons , Parkinson Disease , Parkinsonian Disorders , Substantia Nigra , Zona IncertaABSTRACT
Many seminal advances have been made in human immunodeficiency virus (HIV)/AIDS research over the past four decades. Treatment strategies, such as gene therapy and immunotherapy, are yielding promising results to effectively control HIV infection. Despite this, a cure for HIV/AIDS is not envisioned in the near future. A recently published academic study has raised awareness regarding a promising alternative therapeutic option for HIV/AIDS, referred to as "selective elimination of host cells capable of producing HIV" (SECH). Similar to the "shock and kill strategy," the SECH approach requires the simultaneous administration of drugs targeting key mechanisms in specific cells to efficiently eliminate HIV replication-competent cellular reservoirs. Herein, we comprehensively review the specific mechanisms targeted by the SECH strategy. Briefly, the suggested cocktail of drugs should contain (i) latency reversal agents to promote the latency reversal process in replication-competent reservoir cells, (ii) pro-apoptotic and anti-autophagy drugs to induce death of infected cells through various pathways, and finally (iii) drugs that eliminate new cycles of infection by prevention of HIV attachment to host cells, and by HIV integrase inhibitor drugs. Finally, we discuss three major challenges that are likely to restrict the application of the SECH strategy in HIV/AIDS patients.
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Humans , CD4-Positive T-Lymphocytes , HIV Infections/drug therapy , HIV-1 , Virus LatencyABSTRACT
With the development of the computer simulation technology and the digital simulation technology, the traditional calculation method has been gradually replaced by the digital method to deal the road traffic accident scene and analyse the process. The PC-Crash software simulation system can reconstruct the traffic accidents within 32 vehicles, and the accuracy of reconstruction has been fully verified, which is widely used by the transport police department and the accreditation agency. In this paper, the research of road traffic accident reconstruction using PC-Crash software is reviewed, and the application of road traffic accident reconstruction technology based on PC-Crash software and some existing problems in forensic practice are discussed, which provides reference for the research and identification of road traffic accident simulation and reconstruction and theoretical basis for accident treatment.
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Humans , Accidents, Traffic , Computer Simulation , Models, Theoretical , Police , SoftwareABSTRACT
Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.
Subject(s)
Animals , Humans , Mice , Apoptosis , Arginase , CD11b Antigen , CD4-Positive T-Lymphocytes , Metabolism , Cell Proliferation , Cyclooxygenase 2 , Cyclooxygenase 2 Inhibitors , Gene Expression Regulation , Myeloid Progenitor Cells , Metabolism , Pathology , Nitrobenzenes , Stress Disorders, Traumatic , Drug Therapy , Genetics , Pathology , SulfonamidesABSTRACT
<p><b>OBJECTIVE</b>To study clinical effects of inversive LISS (less invasive stabilization system, LISS) plate combined with steel wire for the treatment of Seinsheimer type V subtrochanteric femoral fractures.</p><p><b>METHODS</b>From January 2009 to Janurary 2012, 11 patients with Seinsheimer type V subtrochanteric femoral fractures were treated with inversive LISS plate combined with steel wire, including 8 males and 3 females, ranging in age from 35 to 61 years old (averaged, 51 years old). Causes of injury: 8 patients caused by accident and 3 patients caused by falling down. According to X-ray examination, all the patients were type V subtrochanteric femoral fractures and were treated with LISS plate combined with steel wire. Merle d'Aubigne scores were used to evaluate therapeutic effects on hip.</p><p><b>RESULTS</b>The mean period of follow-up was 16 months (ranged, 12 to 28 months). All the patients obtained bone union for average 3.8 months (ranged from 3 to 4.3 months). There were no complications such as deep infection,deep vein thrombosis, pulmonary embolism and bone nonunion. According to Merle d'Aubigne scores, 7 patients got an excellent result and 4 good.</p><p><b>CONCLUSION</b>Inversive LISS plate combined with steel wire for the treatment of Seinsheimer type V subtrochanteric femoral fractures is satisfactory, Which can provide stable fixation for fracture end and provide an ideal internal fixation method for treatment of this kind of fractures in the future.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Bone Plates , Bone Wires , Fracture Fixation, Internal , Methods , Hip Fractures , General SurgeryABSTRACT
Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.
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Posterior lumbopelvic fixation with iliac screws is the most commonly used method for unstable spinopelvic injuries. It has certain limitations including inability to use distraction along the spinopelvic rod as an indirect reduction maneuver, need for complex 3-dimensional rod contouring and complications such as hardware prominence and soft tissue coverage. In the present case report, we described a surgical technique of lumbopelvic fixation with sacral alar screws for traumatic spinopelvic instability resulted from a unilateral Denis-III comminuted sacral fracture and the L5 burst fracture. On the opposite side of the sacral fracture, caudal screws were implanted into the pedicle of the S1, whereas on the side of sacral fracture, two sacral alar screws were placed parallel to the superior sacral endplate as well as the plane of sacroiliac joint. In addition, horizontal stabilization was conducted with cross-link connections to maintain the longitudinal traction. For sacral fracture associated with traumatic spinopelvic instability, this modified lumbopelvic fixation technique using sacral alar screws makes longitudinal reduction easier, requires less rod contouring, and reduces hardware prominence without compromising the stability.
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Humans , Bone Screws , Fracture Fixation, Internal , Fractures, Bone , General Surgery , Fractures, Comminuted , Sacrum , General Surgery , Spinal Fractures , General SurgeryABSTRACT
The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified.The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension,and mixed homogeneously into 1.2% alginate gel.Single-cell alginate gel was cultured with serum-free DMEM/F12 medium.Epirubicin(0.8 μg/mL)was added to the medium to enrich CSCs.After cultured conventionally for 7 to 10 days,most of cells suspended in alginate gel were killed by epirubicin.But few cells survived and some single-cell cloning spheres formed.Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation.Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicity in vivo.The results showed that some cells positive for Oct3/4(TRITC)and Nanog(TRITC)were found in single-cell cloning spheres,and most of positive cells were concentrated in the core of sphere.Cells from spheres could form osteosarcoma in the body of mice.It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes(Oct3/4 and Nanog),resisting anti-cancer drugs,and tumorigenicity in vivo.To sum up,it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.
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Interleukin-5 (IL-5) involves in the development of airway inflammation and hyperresponsiveness through activation of eosinophils. Thus, inhibition of IL-5 expression seems to be an attractive approach for asthma therapy. In this study, an antisense IL-5 gene transferred by recombinant adeno-associated virus (asIL-5) was constructed to transfect murine allergic asthma model. Our results showed that asIL-5 efficiently inhibited the IL-5 mRNA expression and significantly attenuated the inflammation in lung tissues. Significant decreasing of eosino-phils and inflammatory cells were found in peripheral blood and bronchoalveolar lavage fluid (BALF). In addition, significant inhibition of airway hyperresponsiveness (AHR) was also found in the mice treated with asIL-5. These observations demonstrate that antisense oligonucleotid against IL-5 delivered by adeno-associated virus system is possibly an efficacious therapeutic strategy for allergic asthma and other eosinophil-related disorders.
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The recombined adenovirus DNA was transfected into 293 cells for packing and amplification of replication-deficient Ad-CMV-E6/E7, Ad-K14 -E6/E7 virus was purified by CsCl density gradient centrifugation , recombined adenovirus Ad-CMV-E6/E7, Ad-K14 -E6/E7 were used as experimental group, while pAd-CMV and pAdtrack-K14 were used as control group. Four of them were injected through one main vein of nude mice tail respectively. These mice were then treated with 0.05 mg 17beta-estradiol over 12 weeks. Mice were anaesthesiaed with 2.5% Avertint and the vagina, mammary gland, ovaries and uterus were dissected and fixed in 3.75% paraformaldehyde overnight at 4 degrees C. Paraffin-embedded sections, HE staining and identification of P53 and Bcl-2 protein via immunohistochemistry were performed. The expression of E6/E7 was verified by RT-PCR in different tissue of nude mice. HE staining showed evident hyperplasy in cervix-uterus transformation zone of experimental group 2. The expression of mutant P53 and Bcl-2 were higher than control group via immunohistochemical S-P method in uterus stroma-cell. Western blotting also showed that E6 protein was expressed. The expression of E6/E7 was higher than control group by human cytokeratin promoter 14 and hyperlasy changes were detected in epithelial tissue of cervix-uterus transformation zone.
Subject(s)
Animals , Female , Humans , Mice , Adenoviridae , Genetics , Blotting, Western , Cell Line , Genital Diseases, Female , Pathology , Virology , Genitalia, Female , Pathology , Virology , Immunohistochemistry , Mammary Glands, Animal , Metabolism , Pathology , Mice, Nude , Oncogene Proteins, Viral , Genetics , Metabolism , Ovary , Metabolism , Pathology , Papillomaviridae , Metabolism , Physiology , Papillomavirus E7 Proteins , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Repressor Proteins , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tumor Suppressor Protein p53 , Metabolism , Uterus , Metabolism , Pathology , Vagina , Metabolism , PathologyABSTRACT
In this study, we used an adenoviral vector -melanoma differentiation-associated gene-7 [Ad-mda7] to examine the effect of the ectopic production of MDA-7/IL-24 on cell migration and invasion by human cervical cancer cells. The study took place in the Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing, China, between April 2006 and November 2006. The change of metastasis of cervical cancer cells [CaSki] cells were detected by Cell Migration Assay and Cell Invasion Assay after treated with Ad-mda7. The production of proteins associated with cell migration and invasion were detected by western blot. Cervical cancer cells treated in vitro with Ad-mda7 migrated and invaded less than cells treated with phosphate-buffered saline [PBS] or Ad-Luc [vector control]. Melanoma differentiation-associated gene-7 /IL-24 inhibited migration and invasion by down-regulating the production of matrix metalloproteinase-2 [MMP-2] and by up-regulating the production of p38 mitogen-activated protein kinase. relative to PBS and Ad-Luc. These results show that MDA-7/IL-24 inhibits invasion and migration by cervical cancer cells by down- or up-regulating proteins associated with these processes, resulting in reduced metastasis. Thus, Ad-mda7 should be considered a therapeutic agent that can inhibit primary tumor growth and prevent metastasis
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<p><b>OBJECTIVE</b>To investigate the clinical manifestations, treatment, complications, and prognosis of patients with multiple injuries.</p><p><b>METHOD</b>The clinical data, including the causes of injury, treatment, complications, causes of death, and mortality rate, of 4519 patients were retrospectively analyzed.</p><p><b>RESULTS</b>The major causes of injury were road traffic injury (2410 cases, 53.33%), violence injury (747 cases, 16.53%), and high falling injury (575 cases, 12.72%). The main involved positions included head (2247 cases, 18.71%), abdominal region and pelvis (2118 cases, 17.64%), and thoracic region (1853 cases, 15.43%). The major complications were shock (1497 cases, 33.13%). The main cause of death was sepsis with multiple organ dysfunction syndrome/failure (28 cases, 82.35%) after multiple injuries, significant higher than other causes in the same period (P<0.01).</p><p><b>CONCLUSIONS</b>The multiple injuries have various causes of disease, and were complicated with their diverse clinical manifestations, numerous complications, and high mortalities. Further research on the integrated rescue mortality is required.</p>
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Humans , Emergency Medical Services , Multiple Trauma , Mortality , Therapeutics , Retrospective StudiesABSTRACT
<p><b>OBJECTIVE</b>To evaluate the value of serum prostate specific antigen (PSA), free PSA (FPSA) and PSA density (PSAD) in early diagnosis of prostatic cancer.</p><p><b>METHODS</b>Sixty-eight patients with benign prostate hyperplasia (BPH), 28 with prostatic intraepithelial neoplasia (PIN) without canceration, and 32 with prostatic cancer, all diagnosed by prostatic biopsy, were enrolled in this study. Serum PSA and FPSA were measured and FPSA/PSA ratio and PSAD calculated for each patient, and the data analyzed to explore the association of these indices with prostatic cancer.</p><p><b>RESULTS</b>Serum PSA level and PSAD were markedly different between the cancer patients and non-cancer patients (P<0.001 and P<0.01, respectively). FPSA/PSA ratio also differed between them (P<0.05). The same results were also found between BPH and cancer patients. Significant difference was noted in serum PSA and PSAD between PIN and cancer patients (P<0.01), but not in FPSA/PSA ratio (P>0.05). No marked difference was observed in serum PSA, FPSA/PSA ratio and PSAD between BPH and PIN patients.</p><p><b>CONCLUSION</b>Serum PSA provides a very important clue for early diagnosis of prostatic cancer, and more accurate diagnosis can be obtained by considering also FPSA/PSA ratio. PSAD may also assist in the early diagnosis of prostatic cancer.</p>
Subject(s)
Aged , Aged, 80 and over , Humans , Male , Middle Aged , Biomarkers, Tumor , Blood , Early Diagnosis , Predictive Value of Tests , Prostate-Specific Antigen , Blood , Prostatic Neoplasms , Blood , DiagnosisABSTRACT
<p><b>OBJECTIVE</b>To analyze the transcription activation and possible regulation mechanism of human X-box binding protein 1(XBP1)gene 5'upstream DNA sequence in different cell lines.</p><p><b>METHODS</b>Six kinds of XBP1 promoter deletion mutants were cloned into pGEM-Teasy vector, which included XBP1 gene 5' upstream -1039 to 66 bp,-859 to 66 bp,-623 to 66 bp,-351 to 66 bp,-227 to 66 bp,-227 to -45 bp respectively. Every deletion mutant sequence was cut from Teasy-XBP1p by KpnI and Xho I, and subcloned into pCAT3-Basic to produce a set of constructs termed as p1-XBP1p, p2-XBP1p, p3-XBP1p, p4-XBP1p, p5-XBP1p, p6-XBP1p, respectively. The transcription activity of each construct was detected after transiently transfecting K562, HepG2,NIH-3T3 and L0(2)cell with FuGENE 6 transfection reagent. Cells transfected by pCAT3-Basic or pCAT3-Promoter were used as negative and positive controls. The activity of chloramphenicol acetyltransferase(CAT), which reflects the transcription activation of the XBP1 gene promoter, was detected by ELISA after 48 hours of transfection.</p><p><b>RESULTS</b>The reporter vectors of six kinds of XBP1 promoter deletion mutants were successfully constructed, as confirmed by restriction enzyme digestion and sequencing. The activities of p4-XBP1p and p5-XBP1p were higher than the other deletion mutants in K562 and HepG2. And the activity of p5-XBP1p was the highest in HepG2. There was no activity detected from any transfected NIH-3T3.</p><p><b>CONCLUSION</b>The XBP1 gene promoter can transactivate its downstream gene to transcription. The core sequence of XBP1 promoter was implied between -227 bp and 66 bp. This sequence was connected with the transcriptional activity of XBP1 promoter closely. Its transcription activity varies with different cell lines. XBP1 promoter might drive gene expression with cell-type specificity.</p>
Subject(s)
Animals , Humans , Mice , 3T3 Cells , 5' Flanking Region , Genetics , Base Sequence , Cell Line , Chloramphenicol O-Acetyltransferase , Metabolism , DNA , DNA-Binding Proteins , Genetics , Gene Deletion , Gene Expression Regulation , Physiology , Genes, Reporter , K562 Cells , Molecular Sequence Data , Nuclear Proteins , Genetics , Promoter Regions, Genetic , Genetics , Regulatory Factor X Transcription Factors , Transcription Factors , Transcription, Genetic , Physiology , Transcriptional Activation , Transfection , Tumor Cells, Cultured , X-Box Binding Protein 1ABSTRACT
<p><b>OBJECTIVES</b>To investigate the regulating effect of HBV pre-S2 protein on iNOS gene promoter and the molecular biological mechanisms of pre-S2 protein in HBV pathogenicity.</p><p><b>METHODS</b>Polymerase chain reaction (PCR) technique was employed to amplify the sequence of iNOS promoter and 3 deletion mutants using HepG2 genomic DNA as the template, and the products were cloned into the pGEM-T vector. The iNOS gene and 3 deletion mutants were cut from T- iNOS by Kpn I and Xho I, and then cloned into pCAT3-Basic. The resulting vectors were named p1-iNOSp, p2-iNOSp, p3-iNOSp, and p4-iNOSp. Each of the reporter vectors was transfected into the HepG2 cell line and cotransfected into HepG2 cells with pcDNA3.1(-)-pre-S2 by FuGENE 6 transfection reagents. The HepG2 cells transfected with pCAT3-Basic were used as a negative control. The activity of CAT in HepG2 cells transfected was detected by an ELISA kit 48 hours after the transfection, which reflected the regulating effect of HBV pre-S2 protein on iNOS gene promoter activity.</p><p><b>RESULTS</b>The expressive vector pcDNA3.1(-)-pre-S2 and report vector pCAT3-iNOSp were constructed and confirmed by restriction enzyme digestion and sequencing. The expression of pcDNA3.1(-)-pre-S2 in HepG2 cells could down-regulate the activity of p1-iNOSp, p3-iNOSp, and the inhibition rate was 54.7% and 79.5%, respectively. The expression of pcDNA3.1(-)-pre-S2 in HepG2 cells had no regulatory effects on p2-iNOSp and p4-iNOSp.</p><p><b>CONCLUSION</b>It is suggested that HBV pre-S2 protein can down-regulate iNOS gene promoter.</p>
Subject(s)
Humans , Down-Regulation , Hepatitis B Surface Antigens , Genetics , Nitric Oxide Synthase Type II , Genetics , Promoter Regions, Genetic , Genetics , Protein Precursors , Genetics , Transcriptional Activation , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To find sensitive and specific micro-metastic markers for prostate cancer.</p><p><b>METHODS</b>Using nested reverse transcription-PCR, we examined the expression of PSA, hK2 and PSMA mRNA in peripheral blood mononuclear cells of 51 patients with prostate cancer, 33 patients with benign prostate hyperplasia (BPH) and 32 normal young people.</p><p><b>RESULTS</b>The expression rates of PSA, hK2 and PSMA mRNA were 52.9%, 43.1% and 64.7%, respectively in prostate cancer group, and 6.2%, 7.7% and 4.6%, respectively in control group (BPH patients and normal young people) with statistical significance (P < 0.01). Although the expression rate of PSA and hK2 mRNA increased with cancer progression, there was no statistical significance among patients in different stages. The expression rate of PSMA mRNA was higher than that of PSA and hK2 mRNA in each clinical stage.</p><p><b>CONCLUSION</b>PSMA mRNA expression detected by nested RT-PCR is of greater value for the diagnosis, therapy choice and prognostic evaluation of prostate cancer patients.</p>
Subject(s)
Aged , Humans , Male , Middle Aged , Antigens, Surface , Blood , Biomarkers, Tumor , Blood , Glutamate Carboxypeptidase II , Blood , Neoplasm Invasiveness , Neoplasm Staging , Prostate-Specific Antigen , Blood , Prostatic Hyperplasia , Blood , Pathology , Prostatic Neoplasms , Blood , Pathology , Tissue Kallikreins , BloodABSTRACT
Objective To investigate the regulating effect of hepatitis B virus(HBV)SPⅠbinding protein 1(SBP1)on inducible nitric-oxide synthase(iNOS)gene promoter activity.Methods Polymerase chain reaction(PCR)technique was employed to amplify the coding sequence of iNOS promoter DNA by using HepG2 genomic DNA as template,and 3 deletion mutants were amplified. The products were cloned into pGEM-T vector,respectively.The iNOS gene and 3 deletion mutants were cut from T-iNOS by KpnⅠand XhoⅠ,and then were cloned into pCAT3-Basic.The construc- ted vectors were named as p1-iNOSp,p2-iNOSp,p3-iNOSp and p4-iNOSp,respectively.Each of the vectors containing different iNOSp DNA fragments was transfected into the HepG2 cell line and cotransfected into HepG2 cells with pcDNA3.1(-)-SBP1 by FuGENE 6 transfection reagents.The HepG2 cells transfected with pCAT3-Basic were used as negative control.The activity of chloram- phenicol acetyltransferase(CAT)in transfected HepG2 cells was detected by an enzyme-linked immu- nosorbent assay(ELISA)kit after 48 hours,which would reflect the regulating effect of SBP1 on iNOS gene promoter activity.Results The expression vector pcDNA3.1(-)-SBP1 and report vector pCAT3-iNOS were constructed and confirmed by restriction enzyme digestion and sequencing.The expression of pcDNA3.1(-)-SBP1 in HepG2 cells up-regulated the activity of p1-iNOSp and down- regulated the activity of p3-iNOSp.The inhibiting rate was 31.3%.Conclusions It is suggested that SBP1 can regulate iNOS gene promoter bidirectionally by influencing the binding sites of nuclear factor (NF)-IL6,A activator domain binding site and NF-?B in iNOS gene promoter.