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Objective:To analyze the damage in hippocampal tissues of mice after whole-body irradiation with high- or low-dose ionizing radiation and to investigate the roles of microglia/macrophages polarization in the injury.Methods:C57BL/6 mice were randomly divided into three groups: sham irradiation group, low-dose group (0.05 Gy) and high-dose group (7 Gy). Low- and high-dose groups were respectively treated by whole-body irradiation with single dose of 60Co γ-rays. Hippocampal tissues of the mice were collected at 6 h, 1 d, 3 d and 7 d after irradiation. The morphology, structure and apoptosis of neurons were detected by HE staining, Nissl staining and Tunnel staining, respectively. RT-PCR and immunofluorescence assay were performed to detect the expression of M1 and M2 microglial markers at mRNA and protein levels in hippocampus tissues. The cognitive and emotional behaviors of mice were evaluated one month after the irradiation by Morris water maze, open field test, elevated plus maze and tail suspension test. Results:There were morphological and structural changes in the nerve cells in the hippocampus region of mice after irradiation, accompanied by apoptosis. Acute injuries occurred at 6 h after radiation, alleviated at 1 d and 3 d, and persisted at 7 d in a dose-dependent manner. The results of immunofluorescence staining and confocal imaging analysis showed that compared with the sham irradiation group, the high-dose group showed increased number of microglia, down-regulated expression of M1 microglial markers and up-regulated expression of M2 microglial markers in the hippocampus at 6 h and 1 d after radiation, while M2 microglial markers decreased at 3 d and 7 d after irradiation. PCR results showed that the expression of M1 and M2 microglial markers at mRNA level in the irradiation groups increased at 6 h after irradiation, but there was no statistical significance. The expression of related proinflammatory/anti-inflammatory factors was significantly up-regulated. The results of behavioral experiments showed that compared with the sham irradiation group, there was no statistical difference in cognitive or emotional behaviors at one month after irradiation.Conclusions:60Co γ-rays could damage mouse hippocampal tissues and result in the overexpression and different polarization patterns of microglia/macrophages in mice.
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Autophagy is a highly conserved intracellular catabolic process used to degrade cytoplasmic components. In recent years, it has attracted much attention because of its importance in the pathogenesis and targeted therapy of acute and chronic kidney disease. Autophagy plays an important role in maintaining renal homeostasis under physiological and pathological conditions. The study of conditional autophagy related gene knockout specific to various renal cells has gradually revealed the role of autophagy in renal diseases. Recent studies have found that autophagy deficiency may play a key role in different pathological states of the kidney. Activated autophagy shows cytoprotective function in both glomerulus and renal tubulointerstitium, suggesting that the up regulation of autophagy may become a potential therapeutic strategy. However, there is also contrary evidence that autophagy may be harmful, which poses a great challenge to the development of therapeutic strategies for up-regulated autophagy.
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Purpose@#Vascular calcification (VC) is a common complication of end-stage renal disease (ESRD). This study aimed to examine changes in the expression of miR-21-5p in ESRD patients with VC and to explore its clinical value in predicting the occurrence and progression of uremic VC. @*Materials and Methods@#120 ESRD patients were divided into patients without VC group (n=38) and patients with VC group (n=82). All patients were followed up for 2 years to evaluate VC progression. qRT-PCR was used to detect serum miR-21-5p levels.Receiver operating characteristic curves were constructed to assess diagnostic value. Kaplan-Meier and log-rank methods were utilized to calculate associations between VC progression and risk factors. @*Results@#Serum miR-21-5p levels were significantly higher in ESRD patients with VC than in those without VC and increased progressively with increasing disease severity. Serum miR-21-5p levels were able to distinguish patients with VC from those without VC, with an area under the curve value of 0.883, a sensitivity of 81.7%, and a specificity of 84.2%. After 2 years of follow-up, miR-21-5p expression had increased in patients with worse VC severity, compared with those with stable VC severity. Patients with high miR-21-5p levels were more likely to develop more severe VC, indicating an association between miR-21-5p and VC progression (log-rank p=0.002). Multivariable Cox regression analysis suggested that serum miR-21-5p is an independent predictive factor of VC progression in ESRD patients (hazard ratio=2.064, 95% confidence interval=1.225–3.478, p=0.006). @*Conclusion@#miR-21-5p is overexpressed in the serum of ESRD patients with VC. Our results suggest that overexpression of miR-21-5p is closely associated with VC progression.
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Occupational hand-arm vibration diseases(HAVD) is a legitimate occupational disease in China, and the mechanism of its pathogenesis is vibration-induced vascular injury. Once HAVD occurs, it is difficult for the patients to recover and can cause great harm to workers exposed to hand-arm vibration. It is difficult to detect and evaluate the occurrence and progress of the disease at an early stage using existing technology, which is disadvantageous to the early prevention and treatment of the disease. Long noncoding RNAs(lncRNAs) play an important role in regulating the development, growth, and remodeling of blood vessels and other biological processes. This article reviews the role and mechanism of lncRNAs in vascular injury, and provides scientific theoretical basis for early diagnosis and treatment of HAVD.
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OBJECTIVE：To explore the mec hanism of baicalein plat elet aggregation inhibitiory effect and lung tissue protective effect of baicalein in model rats with acute pulmonary embolism. METHODS ：Totally 36 rats were randomly divided into normal control group （n＝6）and modeling group （n＝30）. The acute pulmonary embolism model was established by autologous thrombus replication in modeling group ，and the sham operation of rats in normal control group was carried out. After modeling ， 30 model rats were randomly divided into model control group ，positive drug group （low molecular weight heparin calcium 0.01 mL/kg，subcutaneous injection ），baicalein low-dose ，middle-dose and high-dose groups （25，50，100 mg/kg，intraperitoneal injection），with 6 rats in each group. Normal control group and model control group were intraperitoneally injected constant volume of normal saline ；administration groups were given relevant medicine ，once a day ，for consecutive 7 d. After medication ， platelet aggregation rates of rats after activated with adenosine diphosphate （ADP） and arachidonic acid （AA） and platelet activation index （RPI）were detected ；lung histopathology was observed by HE staining ；serum platelet activation markers granule membrane（CD62P）and lysosomal membrane glycoprotein （CD63），growth differentiation factor- 15（GDF-15）and N-terminal B-type natriuretic peptide （NT-proBNP）were measured by ELISA. The mRNA expression levels of Notch 2，Notch3 and Notch signaling ligand PLL 1，JAG2 were detected by RT-PCR method. The protein expression levels of Notch 2，Notch3，DLL1 and JAG2 in lung tissue were detected by immunohistochemistry and Western blotting assay. RESULTS ：Compared with normal control group，plasma ADP-activated platelet aggregation rate ，AA-activated platelet aggregation rate ，RPI，serum levels of CD 62P， CD63，GDF-15 and NT-proBNP were increased significantly （P＜0.05）. The lung tissue of rats was in a state of severe inflammatory infiltration. mRNA and protein expression levels of Notch 2，Notch3，DLL1 and JAG 2 in lung tissue decreased significantly（P＜0.05）. Compared with model control group ，changes of above indexes of rats were improved significantly in baicalein groups （P＜0.05）. CONCLUSIONS ：Baicalein can reduce platelet aggregation and improve the pathological state of lung tissue in rats with acute pulmonary embolism. Its mechanism 0270） may be related to activating Notch signal pathway.
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Objective Diabetic cardiomyopathy (DCM) is one of the complications of diabetes, which is closely related to the change of miRNA. In this study, we observed the characteristic expression of miR-26b in the tissues of the C57BL/6J mouse and in the heart, adipose tissue and liver of the ob/ob mouse, and investigated the effect of high glucose (Glu) on the expression of miR-26b in H9C2 cardiomyocytes. Methods Using RT-PCR, we measured the levels of miR-26b in the heart, adipose tissue, liver and other tissues of C57BL/6J and ob/ob mice. H9C2 cardiomyocytes were treated with Glu at 5.5, 15, 25 and 35 mmol/L for 0, 24, 48, 72, 96 and 120 hours, followed by detection of the proliferation of cardiomyocytes by CCK-8 and determination of thelevels miR-26b. Results The expression of miR-26b was the highest in the heart of the C57BL/6J mice, significantly higher than in the cardiac and white adipose tissues of the ob/ob mice (P < 0.05). The proliferation of cardiomyocytes was markedly increased in the 15, 25 and 35 mmol/L Glu groups at 24, 48, 72, 96 and 120 hours as compared with that in the 5.5 mmol/L Glu group (P < 0.05), higher in the 25 than in the 15 mmol/L Glu group at 24 hours (0.74±0.02 vs 0.72±0.01, P<0.05), but lower in the 35 than in the 15 mmol/L Glu group at 48 hours (0.92±0.01 vs 0.94±0.01, P<0.05), in the 25 and 35 mmol/L Glu groups at 96 hours (P < 0.05), in the 35 mmol/L Glu group at 120 hours (1.12±0.02 vs 1.19±0.05, P<0.05), in the 35 than in the 25 mmol/L Glu group at 24 and 48 hours (P<0.05). The expression of miR-26b in the H9C2 cardiomyocytes was significantly down-regulated in the 25 and 35 mmol/L Glu groups in comparison with that in the 5.5 mmol/L Glu group (P<0.05), remarkably lower in the 25 mmol/L Glu group at 96 and 120 hours than at 0 hour (P<0.05). Conclusion High glucose can down-regulate the expression of miR-26b in H9C2 cardiomyocytes, which suggests that miR-26b may participate in the pathogenesis of DCM.
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Objective To study the role of dendritic cells in the function of a recombinant protein TFPR 1 as an adjuvant .Methods Bone marrow cells were collected from four-to five-week-old male BALB/c mice under aseptic conditions, and cultured with complete RPMI 1640 containing rmGM-CSF and rmIL-4 for six days.TFPR1 was added on day 6, and cells were incubated for another 24 hours.LPS was used as positive control , while PBS as negative .The morphology of dendritic cells was observed under an optical microscope and laser confocal microscope , cell surface makers (CD40,CD80,CD86 and MHCⅡ)were detected with flow cytometry, and the cytokines in the supernatant were detected with ELISA.Results Compared with negative control ,dendritic cells incubated with TFPR1 for 24 hours were significantly different in morphology as was observed by optical and laser confocal microscopes , but were similar to positive control .Most of the dendritic cells treated with TFPR 1 showed less adherence and became round , whose podosomes became shorter , and even disappeared .Actin distribution changed from two poles of the cell to the membrane .CD40,CD80, CD86 and MHCⅡon the cell surface were up-regulated on stimulation by TFPR1,as was detected by FACS.These results showed that TFPR1 was capable of promoting dendritic cell maturation .ELISA showed dendritic cells treated with TFPR 1 secreted high levels of cytokines(IL-6, IL-8 and TNF-α).Conclusion TFPR1 is capable of promoting dendritic cell maturation , and activating cells to produce cytokines , indicating that dendritic cells can play an important role in the function of TFPR 1 as a novel ad-juvant .
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Objective To understand the knowledge,usage,applicability of the standard and the suggestions on the imple-mentation of Criteria for Control and Elimination of Malaria(GB 26345-2010)among malaria prevention and control staff of disease control and medical institutions at all levels in Yunnan Province,so as to provide the evidence for the implementation and revision of the standard.Methods Malaria prevention and control workers at 192 institutions in 16 prefectures and cities of Yunnan Province were surveyed.The malaria prevention and treatment workers at county-level center for disease control and pre-vention(CDCs),county-level medical institutions and township hospitals in Tengchong City and Yingjiang County of Yunnan Province were investigated on the spot.The knowledge and usage,problems and recommendations encountered in the implemen-tation of the standard were collected.Results Totally 444 questionnaires were collected,of which 428 were valid and the valid rate of questionnaires was 96.4％.Among them,the proportion of those who knew the standard was 86.7％(371/428),and the channel of knowledge acquirement was mainly the education and training,accounting for 50.7％(188/371).The total awareness rate of objective indicators in the standard content was 65.9％(282/428).Among the frequency of utilization,the "occasional use" of this standard was the majority,accounting for 33.6％(144/428).Among the applications,the highest proportion of ap-plying to the "regular malaria prevention and control work" was 59.3％(191/322),and only 19.3％(62/322)applied to the "as-sessment for malaria elimination".In the standard suitability assessment,the proportion of considering the standard to be fully applicable was 91.3％(391/428),and the proportion of considering the standard to be revised was 8.7％(37/428).The agen-cies where the respondents were located have taken corresponding measures to promote the implementation of the standard.A to-tal of 43 proposals for the implementation of the standard were collected,relating to personnel and supporting conditions.Con-clusions Combined with the actual situation,the standard should be strengthened practically.At the same time,the standard learning,training,and publicity should be strengthened to raise the implementation level.Its implementation in Yunnan Prov-ince should be sequentially consolidated and steadily promoted.
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Objective To compare the genetic diversity of imported Plasmodium falciparum by Polyα and TAA87 microsatellite markers in Southeast Asian and African geographical isolates. Methods Ninety-two and 126 filter paper samples from patients infected with P. falciparum from Southeast Asia (Myanmar) and Africa (Ghana) were collected, respectively. Two neutral microsatellite loci, Polyα and TAA87 were amplified by PCR. The length of PCR fragments was detected by capillary electrophoresis. The allele frequency and expected heterozygosity (He) were calculated by Excel 2010 and GenALEx 6.0 software. Results A total of 146 P. falciparum samples were analyzed as single infection samples with a total of 26 alleles in locus Polyα and 12 alleles in locus TAA87. The mean He value of the two loci was 0.86 ± 0.02. Ten alleles in locus Polyα and 8 alleles in locus TAA87 were distributed in Myanmar isolates, with the He values of 0.86 and 0.81 respectively. Fifteen alleles in locus Polyα and 11 in locus TAA87 were detected in Ghana isolates, with the He values of 0.91 and 0.86 respectively. In addition, the haplotype of 174 bp (Polyα) and 113 bp (TAA87) were only detected in Myanmar isolates with more than 17% gene frequency, whereas they were absent in Ghana isolates. Conclusions The two different geographical sources of imported P. falciparum strains have different allele frequencies and haplotypes at the two neutral microsatellite markers, Polyα and TAA87. Therefore, these two microsatellite loci may be considered as the potential molecular marker candidates for identifying P. falciparum strains with different geographical sources.
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Objective To compare the genetic diversity of imported Plasmodium falciparum by Polyα and TAA87 microsatellite markers in Southeast Asian and African geographical isolates. Methods Ninety-two and 126 filter paper samples from patients infected with P. falciparum from Southeast Asia (Myanmar) and Africa (Ghana) were collected, respectively. Two neutral microsatellite loci, Polyα and TAA87 were amplified by PCR. The length of PCR fragments was detected by capillary electrophoresis. The allele frequency and expected heterozygosity (He) were calculated by Excel 2010 and GenALEx 6.0 software. Results A total of 146 P. falciparum samples were analyzed as single infection samples with a total of 26 alleles in locus Polyα and 12 alleles in locus TAA87. The mean He value of the two loci was 0.86 ± 0.02. Ten alleles in locus Polyα and 8 alleles in locus TAA87 were distributed in Myanmar isolates, with the He values of 0.86 and 0.81 respectively. Fifteen alleles in locus Polyα and 11 in locus TAA87 were detected in Ghana isolates, with the He values of 0.91 and 0.86 respectively. In addition, the haplotype of 174 bp (Polyα) and 113 bp (TAA87) were only detected in Myanmar isolates with more than 17% gene frequency, whereas they were absent in Ghana isolates. Conclusions The two different geographical sources of imported P. falciparum strains have different allele frequencies and haplotypes at the two neutral microsatellite markers, Polyα and TAA87. Therefore, these two microsatellite loci may be considered as the potential molecular marker candidates for identifying P. falciparum strains with different geographical sources.
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Objective To evaluate the therapeutic effect of 5-aminolevulinic acid mediated photody-namic therapy(ALA-PDT)in combination with recombinant human interferon alpha 2b ointment on superficial basal cell carcinoma(BCC)in elderly.Methods A total of 143 elderly patients with superficial BCC was ran-domly divided into PDT group(n=72 cases)and the surgical group(n=71 cases).The patients in the two groups were given the ALA-PDT in combination with IFN-α2b and routine surgical treatment respectively .The effica-cy after treatment was observed according to surgical wound area , lesion area , recurrence and security during 1 year follow-up.Results The results displayed the complete remission rate of lesion in the PDT group was sig-nificantly higher than that in the surgical group (81.94%vs.57.75%,P0.05).During 1 year follow-up after treatment,the results showed the rate of recurrence in the PDT group was significantly lower than that in the surgical group (9.72% vs.22. 54%,P<0.05).Moreover,the mean wound area,the lesion area in the PDT group after treatment were also sig-nificantly smaller than that in the surgical group (P<0.05).Conclusion ALA-PDT in combination with IFN is an effective method with lower recurrence and minor trauma in the treatment of superficial basal cell carcino -ma.
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We recently developed a mouse model of hepatitis B virus (HBV) chronic infection by intravenous (i.v.) injection with rAAV8-1. 3HBV to C57BL/6 mice. To define the responses of different mouse strains after injection with rAAV8-1. 3HBV, we intravenously injected rAAV8-1. 3HBV at doses of 4 x10(9) (Viral genome,vg), 4 x 10(10) vg and 4 x 10(11) vg to C57BL/6 and BALB/c mice,respectively, and determined the levels of serum HBV antigen and antibody by ELISA,serum viral DNA by real-time PCR,and HBcAg expression in liver by immunohistochemical staining. For C57BL/6 mouse strain with injection of rAAV8-1. 3HBV at three doses, 100% of the mice carried HBV for more than 8 months. The levels of serum HBsAg and HBeAg, serum viral DNA and HBcAg-positive hepatocytes increased in a rAAV8-1. 3HBV dose-dependent manner. For C57BL/6 mice injected with rAAV8-1. 3HBV at the dose of 4 x 10(11) vg,over 40% of hepatocytes expressed HBcAg and serum viral DNA reached over 10(5) IU/mL. No HBV antibody was detected in sera of C57BL/6 mice. For BALB/c mice with injection of rAAV8-1. 3HBV at three doses, serum HBeAg, serum viral DNA and HBcAg-positive hepatocytes persisted for more than 8 months, but serum HBsAg declined remarkably at 2 weeks after injection. The levels of serum HBeAg and HBcAg-positive hepatocytes in BALB/c mice increased in a rAAV8-1. 3HBV dose-dependent manner. Injection with rAAV8-1. 3HBV at the dose of 4 x 10(11) vg resulted in over 50% of BALB/c mice hepatocytes expressing HBcAg. Serum anti-HBsAg were detected in BALB/c mice with rAAV8-1. 3HBV injection at the dose of 4 x10 (10) vg. In conclusion, both C57BL/6 and BALB/c strains can be developed to chronic HBV infection mouse models by i. v. injection with rAAV8-1. 3HBV at doses of 4 x10(9) - 4 x 10(11) vg and the levels of HBV replication increase in a rAAV8-1. 3HBV dose-dependent manner. In contrast to C57BL/6 strain, the BALB/c mice carry out humoral immunity to HBsAg, but fail to mediate HBV clearance.
Subject(s)
Animals , Humans , Male , Mice , Dependovirus , Genetics , Metabolism , Disease Models, Animal , Genetic Vectors , Genetics , Metabolism , Hepatitis B , Allergy and Immunology , Virology , Hepatitis B Antibodies , Allergy and Immunology , Hepatitis B Surface Antigens , Allergy and Immunology , Hepatitis B e Antigens , Allergy and Immunology , Hepatitis B virus , Genetics , Allergy and Immunology , Physiology , Hepatocytes , Allergy and Immunology , Virology , Mice, Inbred BALB C , Mice, Inbred C57BL , Transduction, Genetic , Virus ReplicationABSTRACT
<p><b>OBJECTIVE</b>To investigate the therapeutic effects of closed reduction and external fixation (plaster or splint) for the treatment of displaced humeral supracondylar fractures in children.</p><p><b>METHODS</b>From March 2007 to September 2009,33 children (15 female and 18 male) with humeral supracondylar fractures treated in our hospital, ranging from 3 to 12 years old with an average of 6.4 years old. All the fractures were extension-type injuries, the flexion injures were excluded in our study. The humeral supracondylar fractures were classified according to Gartland classification. There were 21 Type H and 12 type III. In the initial treatment, all the patients were treated with closed reduction and external immobilization. The blood supply of the damaged upper extremity was evaluated before and after treatment. Clinical assessment was obtained at final follow-up using Flynn criteria, and radiologic assessment was obtained using Baumann and lateral humerocapitellar angles.</p><p><b>RESULTS</b>All the children were treated successfully with closed reduction in the initial time; 24 children maintained limb alignment by external immobilization. Nine patients lost position due to the swelling around the elbow which affected unstable external fixation during the follow-up, 5 of which were treated with a repeated closed reduction and internal fixation with Kirschner wires, 4 of which were treated with traction. Thirty-one patients had a satisfactory outcome and 2 patients had an unsatisfactory outcome according to the Flynn criteria at the latest follows-up.</p><p><b>CONCLUSION</b>Closed reduction and external stabilization is an important method for the treatment of displaced humeral supracondylar fractures in children. Making regular follow-up visits after closed reduction and casting is important for patients to maintain acceptable alignment, avoid complications and diagnose any loss of reduction.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Male , Casts, Surgical , External Fixators , Fracture Fixation, Internal , Humeral Fractures , General Surgery , SplintsABSTRACT
<p><b>OBJECTIVES</b>To construct a stable HCV-producing cell model for anti-HCV drug research.</p><p><b>METHODS</b>The HCV-ribozyme recombinant plasmid pJFH1-Rbz was constructed to generate the exact 5' and 3' ends of HCV genomic RNA by placing two self-cleaving ribozymes at both ends of the HCV JFH-1 cDNA. The plasmid was then transfected into HepG2 cells and the resultant clones were screened with G418. Subsequently, immunofluorescence and Western blot were performed to detect the expression of HCV core protein, HCV RNA level was quantitated by TaqMan real-time PCR method and HCV particles was detected by electron microscopy.</p><p><b>RESULTS</b>HCV core protein was detected in the screened cell clone, and the level of HCV RNA was up to 1000,0000 copies/ml in the culture medium. Electron microscopy showed the viral particles in the culture suspension were approximately 55 nm in diameter. IFN-treating experiment demonstrated that the HCV RNA level decreased with the increasing concentration of IFN alpha.</p><p><b>CONCLUSION</b>We constructed a stable HCV-producing cell model which can be used for anti-HCV drug research.</p>
Subject(s)
Humans , DNA, Complementary , Genome, Viral , Hep G2 Cells , Hepacivirus , Genetics , Plasmids , RNA, Catalytic , Genetics , Transfection , Viral Core Proteins , Genetics , Virion , Virus ReplicationABSTRACT
To develop a HBV infection mouse model by hydrodynamic-based transfection and further to optimize the method of development of HBV infection mouse model. We first developed a construct which contained inverted terminal repeat elements (ITR) of adeno-associated virus (AAV) and 1. 3 copies of HBV genome (ayw subtype). The pAAV-HBV1. 3 DNA was then injected hydrodynamically into the tail veins of C57BL/6 mice in 5 seconds. The virus load in serum and liver was assayed by ELISA and Real-time PCR. The expression of virus antigen and the pathologic changes of liver were analyzed by HE and immunohistochemical staining. Meanwhile, to develop HBV transfected immunosuppressied mouse, mice were injected intraperitoneally triple with 0.2 ml dexamethason (50 mg/kg) every two days before HBV transfection. The levels of HBsAg and HBeAg were assayed by ELISA. Our data showed: (1) HBsAg and HBeAg were positive (100%) in serum and liver of experimental normal mouse at day 10 after HBV transfection, and became negative at day 30 and day 60. Meanwhile the viral load in serum and liver in experimental group was significantly higher than that in control group at day 10, 30 and 60 after HBV transfection (P < 0.01, P < 0.05, respectively). (2) HBsAg and HBeAg in serum in immunosuppressed mouse model were positive until 60 days. In conclusion, a HBV infection mouse model was developed successfully by hydrodynamic-based transfection. By suppressing the immune status of mice injected with dexamethasone, the expression of HBV antigens was extended longer than that in normal adult mice. These models pave a way for HBV research and evaluation of HBV vaccine and drug development.
Subject(s)
Animals , Female , Humans , Mice , Dependovirus , Genetics , Metabolism , Dexamethasone , Allergy and Immunology , Disease Models, Animal , Gene Expression Regulation, Viral , Genetic Vectors , Genetics , Metabolism , Hepatitis B , Allergy and Immunology , Virology , Hepatitis B Antigens , Genetics , Metabolism , Hepatitis B virus , Genetics , Physiology , Immunosuppressive Agents , Allergy and Immunology , Liver , Allergy and Immunology , Virology , Mice, Inbred C57BL , Transfection , MethodsABSTRACT
<p><b>OBJECTIVE</b>To highly express TAT-HBX-EGFP fusion protein and study its distribution in mouse liver.</p><p><b>METHODS</b>TAT-HBX-EGFP recombinant vector was constructed and fusion protein was induced by IPTG and expression in BL21; fusion protein was purified by Ni-NTA argarose, then injected into the peritoneal cavity of the mice. Distribution of fusion protein was observed by immunofluorescence.</p><p><b>RESULTS</b>TAT-HBX-EGFP was highly expression in E. coli; HBX could be induced into mouse liver by TAT.</p><p><b>CONCLUSION</b>HBX protein could be induced into mouse liver by TAT induced peptide.</p>
Subject(s)
Animals , Female , Humans , Male , Mice , Cell Membrane , Genetics , Metabolism , Escherichia coli , Genetics , Metabolism , Gene Expression , Green Fluorescent Proteins , Genetics , Metabolism , Hepatitis B , Metabolism , Virology , Liver , Metabolism , Mice, Inbred ICR , Protein Transport , Recombinant Fusion Proteins , Genetics , Metabolism , Trans-Activators , Genetics , Metabolism , Viral Regulatory and Accessory Proteins , Genetics , Metabolism , tat Gene Products, Human Immunodeficiency Virus , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of K18, Ser-33 and Ser-52 phosphorylated K18 in HBV infected human liver disease and its significance.</p><p><b>METHODS</b>The expression and localization of K18 and Ser-33, Ser-52 phosphorylated K18 in healthy liver tissue, in liver tissues of patients with post-HBV infection cirrhosis and severe chronic hepatitis were detected by histochemistry.</p><p><b>RESULTS</b>K18, Ser-33 and Ser-52 phosphorylated K18 were expressed in normal liver cells, in liver tissues of cirrhosis patients and severe chronic hepatitis cases. The expression of K18 in the liver cells from the 3 different sources had no significant difference in levels. Ser-33 and Ser-52 phosphorylated K18 were expressed in normal liver cells, in liver tissues of cirrhosis patients chronicity HBV hepatitis and severe chronic hepatitis cases. Ser-33 and Ser-52 located around cytoplasmic membrane, diffused into cytoplasm and expressed at a higher levels in cirrhosis and severe chronic hepatitis.</p><p><b>CONCLUSION</b>The expression levels of Ser-33 and Ser-52 phosphorylated K18 increased along with the progression of HBV infected human liver disease. The phosphorylation of K18 could be a marker of progression of HBV infected human liver disease.</p>
Subject(s)
Humans , Hepatitis B , Metabolism , Immunohistochemistry , Keratin-18 , Metabolism , Liver Cirrhosis , Metabolism , Pathology , Virology , Liver Diseases , Metabolism , Pathology , Virology , Phosphorylation , Serine , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To analyze the relationship between Fas ligand (FasL) and gastric cancer extensively.</p><p><b>METHODS</b>The computerized CNKI database, Wangfang database, Weipu database, Springerlink full-text periodical database and ProQuest full-text periodical database were searched. And, the literatures detecting FasL in gastric cancer with immunohistochemical method which have been published during 1990 to 2006 were also reviewed. The research hypothesis and research method of each literature were the same; the diagnosis of case and control were definite; the size of sample was also definite; the primitive data must offer the number of object whose FasL were expressed positively or offered the positive rate. Each study and its quality were also evaluated. A software Review Manager 4.2.10, was used to analyze the data. and to estimate the overall OR and its 95% CI.</p><p><b>RESULTS</b>A total of 49 literatures have been reviewed. According to the selection and washing out criteria, 13 literatures were used in this Meta-analysis. In the research of gastric cancer group and normal gastric mucosa group, 10 literatures were used. Between the two groups the expression of FasL has statistical significance (OR(overall) = 14.88, 95% CI 5.34 - 41.48; P < 0.00001); In the research of different differentiation level, 8 literatures were used. Between the two groups the expression of FasL had no statistical significance (OR(overall) = 1.90, 95% CI 0.68 - 5.28; P = 0.22); In the research of different TNM staging, 5 literatures were used. Between the two groups the expression of FasL had statistical significance (OR(overall) = 2.58, 95% CI 1.05 - 6.32; P = 0.04); In the research of different Lymph node metastasis, 8 literatures were used. Between the two groups the expression of FasL has no statistical significance (OR(overall) = 1.00, 95% CI 0.45 - 2.21; P = 1.00).</p><p><b>CONCLUSION</b>The high expression of FasL is the high risk of gastric cancer, and TNM staging has some associations with gastric cancer.</p>
Subject(s)
Humans , Fas Ligand Protein , Metabolism , Risk Factors , Stomach Neoplasms , Epidemiology , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the value of a back propagation (BP) network in determining the risk factors of uterine myomas.</p><p><b>METHODS</b>Using stratified randomized sampling method, 1260 women were surveyed by questionnaire. 1:2 matched case-control study was conducted in 113 cases of uterine myomas. Neural network tools box of Software MATLAB 6.5 was used to train and simulate BP artificial network. The mean impact value (MIV) for each input variables was analyzed, and was compared with multiple logistic regression analysis and log-linear model for interaction between factors.</p><p><b>RESULTS</b>BP artificial neural analysis showed that the leading risk factors for uterine myomas were delayed menstruation, family history of uterine myomas, cervicitis, menstrual disorder, induced abortion, pelvic inflammatory, oral contraceptive medication, and elytritis, with mean impact value -0.0405, 0.0361, 0.0162, 0.0143, 0.0135, 0.0117, 0.0094, 0.0087, respectively. Both BP artificial neural and logistic regression analysis showed that the sequence of leading risk factors were similar in the whole, but there were some differences observed, induced abortion was proved to be an important cooperation variable through logline model analysis respectively.</p><p><b>CONCLUSION</b>Compared to the conventional statistics method, BP artificial neural network could deal with the interaction between covariables preferably, thus provided a powerful method to risk factor analysis.</p>
Subject(s)
Adult , Female , Humans , Middle Aged , China , Epidemiology , Leiomyoma , Epidemiology , Neural Networks, Computer , Risk Assessment , Risk Factors , Uterine Neoplasms , EpidemiologyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the value of a back propogation (BP) network on prediction of birth defect and to give clues on its prevention.</p><p><b>METHODS</b>Data of birth defect in Shenyang from 1995 to 2005 were used as a training set to predict the prevalence rate of birth defect. Neural network tools box of Software MATLAB 6.5 was used to train and simulate BP Artificial Neural Network.</p><p><b>RESULTS</b>When using data of the year 1995-2003 to predict the prevalence rate of birth defect in 2004-2005, the results showed that: the fitting average error of prevalence rate was 1.34%, RNL was 0.9874, and the prediction of average error was 1.78%. Using data of the year 1995-2005 to predict the prevalence rate of birth defect in 2006-2007, the results showed that: the fitting average error was 0.33%, RNL was 0.9954, the prevalence rates of birth defect in 2006-2007 were 11.00% and 11.29%.</p><p><b>CONCLUSION</b>Compared to the conventional statistics method, BP not only showed better prediction precision, but had no limit to the type or distribution of relevant data, thus providing a powerful method in epidemiological prediction.</p>