摘要
OBJECTIVE@#To evaluate the quality of life of patients with interstitial cystitis/bladder pain syndrome (IC/BPS), to compare the difference between IC/BPS and overactive bladder (OAB) pain syndrome, and to explore the related factors affecting the quality of life of IC/BPS patients.@*METHODS@#The demographic data of female outpatients with IC/BPS in Beijing Hospital and other medical centers in China were collected. The quality of life of the patients was investigated by multi-angle questionnaires and compared with the data of OAB patients. According to the influence degree of quality of life, the patients with IC/BPS were divided into mild-moderate group and severe group.@*RESULTS@#In this study, 109 patients with IC/BPS were included. The average age was (46.4±14.3) years and the average course of disease was (39.4±51.6) months. Compared with the OAB patients, the patients in IC/BPS group had a longer average course of disease (P=0.008), a lower proportion of the patients of first visit for the disease (P < 0.001), a higher score of the American Urological Association symptom index (AUA-SI) (P < 0.001), a lower body mass index (BMI) ratio (P=0.016), and a lower incidence of constipation (P=0.006). IC/BPS had the greatest impact on family life, followed by social activity. The score of IC/BPS related symptoms on family life was significantly higher than that of the OAB group (P=0.003). The top three symptoms of the IC/BPS patients were pain (45%), frequency (28%) and urgency (17%). The score of quality of life in the IC/BPS patients was significantly higher than that in the OAB patients (P < 0.001). Caffeine intake (P=0.034) and constipation (P=0.003) might be the factors influencing the quality of life of the patients with IC/BPS.@*CONCLUSION@#IC/BPS has a great influence on the quality of life of patients. Caffeine intake and constipation may be related factors affecting the quality of life of patients with IC/BPS. Urologists should recommend changes in diet and lifestyle to reduce symptoms and improve the patients' quality of life.
Subject(s)
Adult , Female , Humans , Middle Aged , Cystitis, Interstitial/epidemiology , Pain , Quality of Life , Surveys and Questionnaires , Urinary Bladder, Overactive/epidemiology摘要
BACKGROUND@#Dysuria is one of the main symptoms of genitourinary syndrome of menopause, which causes serious disruption to the normal life of peri-menopausal women. Studies have shown that it is related to decrease of detrusor contractile function, but the exact mechanism is still poorly understood. Previous results have suggested that the sphingosine-1-phosphate (S1P) pathway can regulate detrusor contraction, and this pathway is affected by estrogen in various tissues. However, how estrogen affects this pathway in the detrusor has not been investigated. In this study, we detected changes of the S1P/RhoA/Rho associated kinases (ROCK)/myosin light chain (MLC) pathway in the detrusor of ovariectomized rats in order to explore the underlying mechanism of dysuria during peri-menopause.@*METHODS@#Thirty-six female Sprague-Dawley rats were randomly divided into SHAM (sham operation), OVX (ovariectomy), and E groups (ovariectomy + estrogen), with 12 rats in each group. We obtained bladder detrusor tissues from each group and examined the mRNA and protein levels of the major components of the S1P/RhoA/ROCK/MLC pathway using quantitative real-time polymerase chain reaction and Western blotting, respectively. We also quantified the content of S1P in the detrusor using an enzyme linked immunosorbent assay. Finally, we compared results between the groups with one-way analysis of variance.@*RESULTS@#The components of the S1P pathway and the RhoA/ROCK/MLC pathway of the OVX group were significantly decreased, as compared with SHAM group. The percent decreases of the components in the S1P pathway were as follows: sphingosine kinase 1 (mRNA: 39%, protein: 45%) (both P 0.05). The percent decreases of the components in the RhoA/ROCK/MLC pathway were as follows: ROCK2 (protein: 41%, mRNA: 36%) (both P 0.05). In addition, all of the above-mentioned decreases could be reversed after estrogen supplementation (E group vs. SHAM group) (all P > 0.05).@*CONCLUSION@#In this study, we confirmed that ovariectomy is closely associated with the down-regulation of the S1P/RhoA/ROCK/MLC pathway in the rat detrusor, which may be one mechanism of dysuria caused by decreased contractile function of the female detrusor during peri-menopause.
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Objective: To explore the key genes and potential therapeutic drugs for osteoarthritis (OA) by bioinformatics.Method: The microarray data GSE55235 was downloaded from the data platform of gene expression omnibus (GEO) and the differentially expressed genes were screened by R language software (3.5.0).Then,the differentially expressed genes were subjected to gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway analysis with David online database.The protein-protein interaction was analyzed by String 10.5 online database and visual editing was analyzed by Cytoscape v3.6.1 software.Subnetwork module analysis was utilized by MCODE plugin to screen the core genes in the process of OA.Finally,small molecule drugs with potential treatment for OA were analyzed by connectivity map (CMap) database.Result: A total of 556 differentially expressed genes were screened,among which 252 were up-regulated and 304 were down-regulated.These genes were mainly involved in extracellular matrix (ECM) organization,inflammatory response,cell adhesion,immune response,collagen binding,etc.The analysis of KEGG pathway showed that differential genes were mainly involved in ECM-receptor interaction,phosphatidylinositol 3 kinase-protein kinase B (PI3K/Akt) signaling pathway and osteoclast differentiation.Some genes,such as interleukin-6(IL-6),JUN,vascular endothelial growth factor α(VEGFA),FOS,MYC and early growth response gene-1(EGR-1),activating transcription factor-3(ATF-3),playing critical role in the process of OA were identified by protein-protein interaction.Some potential small molecular drugs for the treatment of OA have also been screened,such as lycorine and anisomycin.Conclusion: The selected key genes may be targets for the diagnosis of OA or potential targets for the treatment of OA,and the selected small molecular drugs can be developed as the key drugs for the treatment of OA.
摘要
Buyang Huanwu Decoction(BHD) has the effect in benefiting Qi and activating blood circulation,and is the representative prescription for benefiting Qi and activating blood. At present,it is used for treatment of early diabetic nephropathy. However,its efficacy and safety remained to be verified. Therefore,this study aims to systematically evaluate the efficacy and safety of Buyang Huanwu Decoction for early-stage diabetic nephropathy. Data of randomized controlled trials(RCTs) of Buyang Huanwu Decoction for earlystage diabetic nephropathy were collected through the retrieval of electronic databases,including PubMed,EMbase,the Cochrane Library,CBM,CNKI,VIP and Wan Fang Data from inception to September 16,2018. Two reviewers independently screened out literatures,extracted data,and assessed the risk of bias. And then Meta-analysis was conducted by Rev Man 5. 3 software. A total of 15 RCTs involving 1 402 patients were included. The results of Meta-analysis indicated that Buyang Huanwu Decoction and conventional treatment group(combination treatment group) were superior to conventional treatment group in reducing 24 h urinary albumin excretion rates(MD =-40. 23,95% CI[-71. 25,-9. 21],P = 0. 01) and total cholesterol(MD =-0. 75,95% CI[-1. 02,-0. 48],P <0. 000 01). The effects of the two groups in reducing serum creatinine were similar(MD =-1. 48,95%CI[-4. 48,1. 53],P = 0. 34).However,the reduction of triglyceride was affected by the course of treatment. The effects were similar in less than or equal to eight weeks(MD =-0. 33,95%CI[-0. 97,0. 31],P = 0. 31),whereas the combination group was superior to the conventional treatment group in 12 weeks(MD =-0. 30,95%CI[-0. 58,-0. 22],P = 0. 03) and more than or equal to 16 weeks(MD =-0. 49,95% CI[-0. 9,-0. 08],P= 0. 02). There were no significant difference in adverse events between the two groups(OR= 1. 38,95%CI[0. 28,6. 8],P = 0. 69). The results showed that combination treatment group has a significantly higher efficacy on early diabetic nephropathy.The above conclusion shall be verified with more high-quality RCTs because of the low quality of the included studies.
Subject(s)
Humans , Diabetic Nephropathies , Drug Therapy , Drugs, Chinese Herbal , Therapeutic Uses , Randomized Controlled Trials as Topic摘要
Objective:To investigate the effect of miR-21 on the proliferation and apoptosis of leukemia cell line K562 and the regulation of PI3K/AKT signaling pathway.Methods:K562 cells were divided into control group,miR-21 NC group and miR-21 interference group,the first group without treatment,the two groups after using cationic liposome transfection of LipofectamineTM2000 miR-21 inhibitor and miR-21 negative control.After transfection 48 h,the expression of miR-21 mRNA in cells was detected by Real-time fluorescence quantitative (qRT-PCR),MTT assay was used to test the effect of miR-21 on cell proliferation,flow cytometry was used to detected the effect of miR-21 on cell cycle apoptosis rate,the protein expression of PI3K,AKT and p-AKT in PI3K/AKT signaling pathway after miR-21 transfection were detected by protein immunoblotting (Western blot).Results:The expression level of miR-21 mRNA and the survival rate of cells in the miR-21 interference group were significantly lower than those in the control group and the miR-21 NC group;compared with control group and miR-21 group NC,flow cytometry detected the cells accounted for proportion of G0/G1 phase in miR-21 interference group increased significantly,the proportion of cells at S phase significantly decreased,and apoptosis rate increased significantly.Western blot test results show that the expression levels of p-AKT in miR-21 inter-ference group were significantly lower than those in the control group and miR-21 NC group,but the expression levels of PI3K and AKT protein were not changed significantly.Conclusion:Down-regulation of miR-21 can inhibit the proliferation and promote apoptosis of leukemia K562 cells,and its mechanism may be related to the inhibition of PI3K/AKT signaling pathway.
摘要
OBJECTIVE@#This work aimed to evaluate the influence of smear layer on the bonding effectiveness and durability of the self-adhesive resin cements to dentin.@*METHODS@#A total of 48 fresh caries-free third molars with exposed dentin surface were divided into two groups. The dentin surfaces were treated using a standard grit diamond bur (group A) or further polished using a fine grit diamond bur (group B) and then bonded with either of the two self-adhesive resin cements, namely, Clearfil SA Cement (CSA, Kuraray) and Multilink Speed (MS, Ivoclar Vivadent). After 24 h or 2-year water storage, a microtensile bond strength test was performed.@*RESULTS@#In group A, the dentin surface was rough, the smear layer was thick, and the dentin tubule orifice detritus showed low embolism value. In group B, the dentin surface roughness decreased, the embolism proportion increased, and the smear layer became thin. The initial bonding strengths of CSA and MS in group A were significantly lower than those in group B (P0.05).@*CONCLUSIONS@#The properties of the smear layer and the types of self-adhesive resin cement used affected the bond strength and durability.
Subject(s)
Humans , Composite Resins , Dental Bonding , Dental Cements , Dentin , Dentin-Bonding Agents , Materials Testing , Resin Cements , Smear Layer , Surface Properties , Tensile Strength摘要
The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites.
Subject(s)
Animals , Female , Male , Animals, Domestic/blood , Antibodies, Fungal/blood , Antibodies, Protozoan/blood , China/epidemiology , Encephalitozoon cuniculi/immunology , Encephalitozoonosis/blood , Rabbits/blood , Seroepidemiologic Studies , Toxoplasma/immunology , Toxoplasmosis, Animal/blood摘要
Pyronaridine and artesunate have been shown to be effective in falciparum malaria treatment. However, pyronaridine is rarely used in Hainan Island clinically, and artesunate is not widely used as a therapeutic agent. Instead, conventional antimalarial drugs, chloroquine and piperaquine, are used, explaining the emergence of chloroquine-resistant Plasmodium falciparum. In this article, we investigated the sensitivity of P. falciparum to antimalarial drugs used in Hainan Island for rational drug therapy. We performed in vivo (28 days) and in vitro tests to determine the sensitivity of P. falciparum to antimalarial drugs. Total 46 patients with falciparum malaria were treated with dihydroartemisinin/piperaquine phosphate (DUO-COTECXIN) and followed up for 28 day. The cure rate was 97.8%. The mean fever clearance time (22.5+/-10.6 hr) and the mean parasite clearance time (27.3+/-12.2 hr) showed no statistical significance with different genders, ages, temperatures, or parasite density (P>0.05). The resistance rates of chloroquine, piperaquine, pyronarididine, and artesunate detected in vitro were 71.9%, 40.6%, 12.5%, and 0%, respectively (Ppiperaquine>pyronarididine>artesunate. The inhibitory dose 50 (IC50) was 3.77x10(-6) mol/L, 2.09x10(-6) mol/L, 0.09x10(-6) mol/L, and 0.05x10(-6) mol/L, and the mean concentrations for complete inhibition (CIMC) of schizont formation were 5.60x10(-6) mol/L, 9.26x10(-6) mol/L, 0.55x10(-6) mol/L, and 0.07x10(-6) mol/L, respectively. Dihydroartemisinin showed a strong therapeutic effect against falciparum malaria with a low toxicity.
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Antimalarials/pharmacology , China , Inhibitory Concentration 50 , Malaria, Falciparum/drug therapy , Parasitic Sensitivity Tests , Plasmodium falciparum/drug effects , Treatment Outcome摘要
Pyronaridine and artesunate have been shown to be effective in falciparum malaria treatment. However, pyronaridine is rarely used in Hainan Island clinically, and artesunate is not widely used as a therapeutic agent. Instead, conventional antimalarial drugs, chloroquine and piperaquine, are used, explaining the emergence of chloroquine-resistant Plasmodium falciparum. In this article, we investigated the sensitivity of P. falciparum to antimalarial drugs used in Hainan Island for rational drug therapy. We performed in vivo (28 days) and in vitro tests to determine the sensitivity of P. falciparum to antimalarial drugs. Total 46 patients with falciparum malaria were treated with dihydroartemisinin/piperaquine phosphate (DUO-COTECXIN) and followed up for 28 day. The cure rate was 97.8%. The mean fever clearance time (22.5+/-10.6 hr) and the mean parasite clearance time (27.3+/-12.2 hr) showed no statistical significance with different genders, ages, temperatures, or parasite density (P>0.05). The resistance rates of chloroquine, piperaquine, pyronarididine, and artesunate detected in vitro were 71.9%, 40.6%, 12.5%, and 0%, respectively (Ppiperaquine>pyronarididine>artesunate. The inhibitory dose 50 (IC50) was 3.77x10(-6) mol/L, 2.09x10(-6) mol/L, 0.09x10(-6) mol/L, and 0.05x10(-6) mol/L, and the mean concentrations for complete inhibition (CIMC) of schizont formation were 5.60x10(-6) mol/L, 9.26x10(-6) mol/L, 0.55x10(-6) mol/L, and 0.07x10(-6) mol/L, respectively. Dihydroartemisinin showed a strong therapeutic effect against falciparum malaria with a low toxicity.
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Antimalarials/pharmacology , China , Inhibitory Concentration 50 , Malaria, Falciparum/drug therapy , Parasitic Sensitivity Tests , Plasmodium falciparum/drug effects , Treatment Outcome摘要
<p><b>OBJECTIVE</b>To explore the polymorphism in circumsporozoite protein of Plasmodium vivax before malaria was eliminated in Hainan island.</p><p><b>METHODS</b>PCR amplification, sequencing, and alignment methodologies were conducted and phylogenetic tree constructed.</p><p><b>RESULTS</b>From all the cases, 19 of them belonged to two types, with 18 as VK210 type and 1 as VK247 type. VK210 type could be divided into seven kinds of subtypes but VK247 had only one type. Ratio of tropical strain with temperate stain in VK210 type was explored between the two stages:control or elimination. Phylogenetic tree was constructed by amino acid sequencing which clearly manifested that VK210 type and VK247 type belonged to different clusters.</p><p><b>CONCLUSION</b>Compared the proportion of two types in the control stage, there was no significant difference seen in the stage of elimination.</p>
Subject(s)
Humans , China , Epidemiology , Genotype , Malaria, Vivax , Epidemiology , Plasmodium vivax , Classification , Genetics , Polymorphism, Genetic , Spores, Protozoan , Genetics摘要
<p><b>OBJECTIVE</b>To evaluated the effect of curing modes and light-cure times on knoop hardness (KH) and microtensile bond strength (µTBS) of dentin adhesives in vitro.</p><p><b>METHODS</b>Twenty molars were made into 80 dentin slices (about 1 mm thick). The dentin slices were prepared with an etch&rinse adhesive A (ONE-STEP PLUS) and a self-etch adhesive B (Clearfil SE Bond), and light-cured respectively under fast mode, i.e.1250 mW/cm(2) light intensity for 10 s, 15 s, 20 s, and ramp mode (soft start curing mode), i.e.initial 0 mW/cm(2) gradually increasing to 1250 mW/cm(2) in first 10 s, then steady for the next 10 s. The prepared dentin slices were kept in dark dry room for 24 h at 37°C, and KH were tested. The other 40 molars were flattened to expose coronal dentin, prepared with adhesives as above. Then the prepared teeth were restored with resin composites incrementally and cured under fast mode. The restored teeth were stored in water for 24 h at 37°C, and slowly sectioned to obtain multiple bonded beams. After 7 d water-storage, the samples received microtensile bond test, and the failure models of beams were observed under a stereomicroscope. Data were analyzed by ANOVA and LSD test (α = 0.05).</p><p><b>RESULTS</b>No statistical difference in KH [(28.20 ± 5.36), (29.13 ± 5.60), (28.13 ± 4.40), (27.06 ± 3.77) MPa] and µTBS [(22.30 ± 5.07), (22.73 ± 6.59), (26.32 ± 6.17), (25.67 ± 4.31) MPa] of adhesive A were found between four curing conditions (fast mode for 10 s, 15 s, 20 s and ramp mode for 20 s) (P > 0.05). In adhesive B, KH of Fast 20 s [(28.23 ± 3.67) MPa] were significantly higher than those of Fast 10 s [(14.15 ± 2.24) MPa] and Fast 15 s [(17.63 ± 2.17) MPa] (P < 0.05). The µTBS of Fast 20 s [(42.52 ± 3.59) MPa] were significantly higher than those of Fast 10 s [(24.21 ± 3.60) MPa], Fast 15 s [(22.25 ± 4.16) MPa] and Ramp 20 s [(31.12 ± 5.40) MPa] (P < 0.05). In Fast 20 s and Ramp 20 s modes, there were no statistical difference in KH of adhesive A and B, while µTBS of adhesive B were higher than that of adhesive A(P < 0.05).</p><p><b>CONCLUSIONS</b>As for different type dentin adhesives, the appropriate curing time in fast mode is different, and ramp mode (soft start curing mode) has no advantage over fast mode.</p>
Subject(s)
Humans , Dentin-Bonding Agents , Hardness , In Vitro Techniques , Light-Curing of Dental Adhesives , Methods , Tensile Strength摘要
In contrast to the gradual reduction in the number of locally transmitted malaria cases in China, the number of imported malaria cases has been increasing since 2008. Here, we report a case of a 39-year-old Chinese man who acquired Plasmodium ovale wallikeri infection while staying in Ghana, West Africa for 6 months in 2012. Microscopic examinations of Giemsa-stained thin and thick blood smears indicated Plasmodium vivax infection. However, the results of rapid diagnostic tests, which were conducted 3 times, were not in agreement with P. vivax. To further check the diagnosis, standard PCR analysis of the small-subunit rRNA gene was conducted, based on which a phylogeny tree was constructed. The results of gene sequencing indicated that this malaria is a variant of P. ovale (P. ovale wallikeri). The infection in this patient was not a new infection, but a relapse of the infection from the one that he had contracted in West Africa.
Subject(s)
Adult , Humans , Male , Azure Stains , Base Sequence , China , DNA, Protozoan/chemistry , DNA, Ribosomal/chemistry , Ghana , Malaria/diagnosis , Molecular Sequence Data , Phylogeny , Plasmodium ovale/classification , Polymerase Chain Reaction , Recurrence , Sequence Analysis, DNA , Travel摘要
To investigate whether HIV-1 infection affects expression of interferon-stimulated gene 15 (ISG15) and determine the antiviral effect of ISG15 in vitro, ISG15 expression at transcription and protein level and supernatant p24 of HIV-1 was detected in various HIV-1 infected or transfected cell lines, respec tively. HIV-1 molecular clone pNL4-3 was used to transfect 293T, TZM-bl and HeLa cells while HIV-1 pseudo-typed virus was used to infect Jurkat, MT-1 and THP-1 cells. After twenty-four hours post infection or transfection, cells were harvested for extraction of total RNAs and subsequently used in real time PCR for quantification of ISG15 transcriptional expression. After forty-eight hours post infection or transfection, cells were harvested for extraction of total proteins to detect ISG15 protein expression. A significant up-regulation of ISG15 at transcription level was observed in HIV-1 infected or transfected cell lines, particulaly in THP-1 and TZM-bl cells. Up-regulation of ISG15 protein was observed only in TZM-bl cell. Cotransfection of ISG15 and HIV-1 indicated that ISG15 inhibited production of HIV-1 progeny virus in a dose and time depend manner in 293T cell but not TZM-bl cell. These results revealed upregulating ISG15 expression in transcriptional level and potential antagonistic mechanism against ISG15 by HIV-1 infection, simultanelusly.
Subject(s)
Humans , Base Sequence , Cell Line , Cytokines , Genetics , Metabolism , HIV Infections , Genetics , Metabolism , Virology , HIV-1 , Physiology , Interferons , Metabolism , Molecular Sequence Data , Ubiquitins , Genetics , Metabolism , Up-Regulation摘要
<p><b>BACKGROUND</b>The Fc receptor associated pathway might improve the immune responses against hepatitis B virus (HBV) as previously described by us. In addition, the Flt3 ligand (FL) has been reported to potentiate antigen presenting cells in vivo and may act as a potential adjuvant to boost antigen-specific immune responses. In this study, the immune efficacies of a set of fusion proteins of HBsAg and Fc and/or FL were evaluated in HBsAg transgenic mice.</p><p><b>METHODS</b>The fusion proteins composed of HBsAg and the Fc domain of murine IgG1 (HBsAg-Fc) and/or the Flt3 ligand, and yeast-derived recombinant HBsAg were used as immunogen to immunize HBsAg transgenic mice, respectively. Serum and liver HBsAg levels, serum anti-HBsAg and cytokine profile, and the activities of alanine aminotransferase (ALT)/AST were investigated after immunization.</p><p><b>RESULTS</b>After six injections, the most pronounced decrease in serum and liver HBsAg levels was observed in the HBsAg-Fc immunized group. In addition, serum Th1 cytokines and ALT/AST activities were highest in this group, indicating an effective induction of a favorable cellular immune response. Interestingly, the fusion protein containing HBsAg-Fc and the Flt3 ligand stimulated an alternative Th1-type immune response featured with high level productions of tumor necrosis factor α (TNF-α) and monocyte chemoabstractant protein 1 (MCP-1), causing a more severe cytotoxicity in hepatocytes while showed less effective in reducing serum HBsAg level.</p><p><b>CONCLUSION</b>HBsAg-Fc is effective in eliciting both the humoral and cellular immune responses against HBsAg in HBsAg transgenic mice, which makes it a potential immunogen for the immunotherapy of chronic hepatitis B.</p>
Subject(s)
Animals , Female , Male , Mice , Chemokine CCL2 , Metabolism , Cytokines , Metabolism , Enzyme-Linked Immunosorbent Assay , Hepatitis B Surface Antigens , Genetics , Allergy and Immunology , Metabolism , Immunity, Cellular , Allergy and Immunology , Immunity, Humoral , Allergy and Immunology , Mice, Inbred C57BL , Mice, Transgenic , Receptors, Fc , Genetics , Allergy and Immunology , Metabolism , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Metabolism , Tumor Necrosis Factor-alpha , Metabolism摘要
To develop polyclonal antibodies against predicted B cell epitopes in HIV-1 accessory protein Vpr, the prepared consensus Vpr amino acid sequence was used to predict potential B cell epitopes by online softwares (ABCpred and Bcepred), the synthesized polypeptides of B-cell epitopes were subsequently conjugated with keyhole limpet hemocyanin (KLH) and then used to immunize rabbits. The antibody titers were determined by ELISA, and antibody specifity was analyzed by Western-Blotting and immunoprecipitation, respectively. Amino acid residues 3-19 (N) and 82-95 (C) of Vpr were predicted as the potential B cell epitopes. After inoculation of the conjugation of synthesized peptide to KLH, the antibody titers in rabbit sera against N and C peptides reached more than 1:100000 by ELISA. Western-Blotting analysis showed that the polyclonal antibodies reacted with both wild Vpr and fusion protein of GFP with Vpr, no matter Vpr was derived from HIV-1 B subtype or CRF07_BC recombinant form; Immunoprecipitation analysis showed similar reactions to Western-Blotting results. Two B cell epitopes of Vpr were successfully predicted by Bio-informatics methods and polyclonal antibodies against those peptides could be successfully prepared.
Subject(s)
Animals , Humans , Rabbits , Antibodies, Viral , Blood , Allergy and Immunology , Cell Line , Enzyme-Linked Immunosorbent Assay , HIV Infections , Blood , Allergy and Immunology , Virology , HIV-1 , Genetics , Allergy and Immunology , Peptides , Genetics , Allergy and Immunology , vpr Gene Products, Human Immunodeficiency Virus , Genetics , Allergy and Immunology摘要
<p><b>BACKGROUND</b>Although it was widely accepted that full-length HIV genome sequences is important in studying virus genetic evolution and variation as well as developing vaccine candidate, to directly sequencing HIV-1 genome of virion RNA remains as a challenge worldwide. Up to date, no published genomic sequences from virion RNA are available for Chinese prevalent HIV-1 strains due to the absence of specialized protocol and appropriate lab equipments. In this study we developed a straightforward approach for amplifying and sequencing HIV virion RNA from plasma by modifying published protocols and further confirmed it is suitable to process Chinese samples.</p><p><b>METHODS</b>The methods for viral RNA extraction and gene amplification was modified and optimized as could be widely used in most Chinese labs. Gene alignment of Chinese HIV-1 strains was employed for designing specialized primer sets for Thai-B and BC recombinant strains. Based on comprehensively consideration of high variable gene region and recombinant breakpoints in BC recombinant strains, a three-amplicon strategy (including 4.3-kb gag-pol, 2.9-kb pol-env and 2.7-kb env-nef) was developed. In addition, one amplicon (9 kb near full-length genome) was also used in 32 samples with varied viral loads. All amplicons were directly sequenced by DNA automated sequencer.</p><p><b>RESULTS</b>Twenty-five percent (8/32) amplification efficiency was achieved by the one-amplicon strategy and 65.6% (21/32) by three-amplicon strategy. For one amplicon strategy, none of complete near full-length genome sequences was obtained by DNA sequencing. For three-amplicon strategy, 75% sequences were achieved in DNA sequencing. Amplification efficiency but not sequencing efficiency was closely associated with viral loads.</p><p><b>CONCLUSION</b>Three-amplicon strategy covering all encoding regions of HIV-1 is suitable for Thai-B and BC recombinant strains and could be potentially employed in less-well equipped Chinese labs.</p>
Subject(s)
Genome, Viral , Genetics , HIV-1 , Genetics , RNA, Viral , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Virion , Genetics摘要
<p><b>OBJECTIVE</b>To investigate the feasibility of prevention and treatment of early scar through observing the effect of feeding immunosuppressive drug cyclophosphamide on rabbit ears hypertrophic scar tissue in early stage.</p><p><b>METHODS</b>Thirty-two Rabbit ears were used to establish animal models for hypertrophic scar and randomly divided into four groups: group of distilled water (A), group of cyclophosphamide 5 mg x kg(-1) x d(-1) (B), group of 10 mg x kg(-1) x d(-1) (C), group of 30 mg x kg(-1) x d(-1) (D). Before animal models were built and after administration for 14 days, 28 days, leukocytes and lymphocytes were detected. After 28 days, specimens were harvested and underwent HE staining and VG staining in order to assess HI, NA, AA value changes. The data (HI, NA, AA) from each group were compared by analysis of variance, and the variance for the rank sum test when missing.</p><p><b>RESULTS</b>On the 14th day, the number of leukocytes in group A, B, C, D were (8.62 +/- 0.58) x 10(9)/L, (4.48 +/- 0.41) x 10(9)/L, (2.7 +/- 0.26) x 10(9)/L, (1.33 +/- 0.27) x 10(9)/L; the number of lymphocytes in group A, B, C, D were (3.11 +/- 0.21) x 10(9)/L, (1.67 +/- 0.16) x 10(9)/L, (0.42 +/- 0.10) x 10(9)/L, (0.40 +/- 0.09) x 10(9)/L. On the 28th day, the number of leukocytes in group A, B, C, D was (8.63 +/- 0.53) x 10(9)/L, (5.10 +/- 0.27) x 10(9)/L, (3.10 +/- 0.26) x 10(9)/L, (1.98 +/- 0.20) x 10(9)/L; the number of lymphocytes A, B, C, D was (3.06 +/- 0.16) x 10(9)/L, (2.08 +/- 0.14) x 10(9)/L, (0.96 +/- 0.19) x 10(9)/L, (0.14 +/- 0.07) x 10(9)/L. On the 14th day and 28th day, the number of leukocytes and lymphocytes in experimental groups was reduced, showing a negative relation with cyclophosphamide dose (P < 0.05). The HI in group of A, B, C, D was 3.02 +/- 0.24, 2.59 +/- 0.43, 2.06 +/- 0.19, 1.63 +/- 0.11; the AA was 40.49 +/- 2. 07, 35.29 +/- 1.99, 28.36 +/- 1.87, 24.99 +/- 1.82; the NA was 4570.5 +/- 259.3, 4222.5 +/- 199.6, 3540.3 +/- 170.3, 3341.4 +/- 228.8. The difference in HI, AA, NA between control group and any of the experimental groups was statistically significant (P < 0.01). Each group, with the dose increased, except NA content of group C and D, the HI, AA, NA was more smaller, negative correlation, the difference was statistically significant (P < 0.05).</p><p><b>CONCLUSIONS</b>Feeding cyclophosphamide can inhibit leukocytes and lymphocytes number, so as to inhibit the proliferative activity of hypertrophic scar. It has significant effect on prevention of hypertrophic scar on rabbit ears in early stage.</p>
Subject(s)
Animals , Female , Male , Rabbits , Cicatrix, Hypertrophic , Drug Therapy , Cyclophosphamide , Pharmacology , Ear , Pathology , Leukocyte Count , Leukocytes , Lymphocyte Count , Lymphocytes摘要
<p><b>OBJECTIVE</b>To evaluate the effect of different silane couplers on bond strength and durability of two machinable glass ceramics to resin cement.</p><p><b>METHODS</b>Two machinable glass ceramics (A and B) were silanized by three silane couplers (A, B, C), and were bonded with a resin cement (G-CEM) to form micro-shear test specimens of six groups. The specimens of each group were subdivided into two subgroups, and their micro-shear bond strength was measured before and after 10000 thermal cycles. Bond strength data were analyzed by two-way ANOVA.</p><p><b>RESULTS</b>Before thermal cycles, the bond strength of ceramic A treated by silane coupler A was lower than that of ceramic B (P = 0.002). The bond strength of ceramic A treated by silane coupler C was significantly higher than that treated by silane coupler A and B (P = 0.014, P = 0.019). 10 000 thermal cycles obviously decreased the bond strength of all groups except the group of ceramic A treated by silane coupler B, and no significant difference was found between three silane coupler with either of two ceramic. However the bond strength of ceramic B treated by silane coupler B and C was significantly higher than that of ceramic A (P = 0.003, P = 0.027).</p><p><b>CONCLUSION</b>As well as the types of silane coupler, the type of ceramic could affect their bond strength and durability to resin cement.</p>
Subject(s)
Ceramics , Materials Testing , Resin Cements , Silanes摘要
<p><b>OBJECTIVE</b>To evaluate the effect of Al2O3 particles sandblasting on the surface roughness, element composition and resin bond durability of zirconia ceramic.</p><p><b>METHODS</b>Sixty 2.5 mm thick computer aided design and computer aided manufacture (CAD/CAM) zirconia ceramic (Vita Inceram YZ) plates were fired, polished and cleaned. Half of polished ceramic plates was sandblasted with 50 µm alumina particles at 0.3 MPa for 20 s. The surface roughness of polished and sandblasted ceramic surface were measured by 3D-laser scanning microscope, and the surface element weight and atom ratio of the ceramic surface were measured by energy disperse spectroscopy (EDS). Then polished and sandblasted ceramic plates were randomized into six groups. In Group 1 and 2 the polished and sandblasted ceramic plates were bonded irrespectively with conventional resin cement (DUOLINK). In Group 3 and 4 the ceramic plates were bonded with resin cement containing MDP (Panavia F), In Group 5 and 6 the specimens were pretreated with silane coupler acitivated by MDP (Clearfil Ceramic Primer), then bond with Panavia F. The specimens of each test group were then divided into two subgroups, and to received shear test after 0 and 10 000 time thermal cycle. The data was analyzed by one-way ANOVA and independent t test.</p><p><b>RESULTS</b>Comparing with polishing, sandblasting reduced the oxygen atom and weight ratio of zirconia ceramic surface (P < 0.001), and increased the zirconium atom and weight ratio (P < 0.001), meanwhile increased the surface roughness (P < 0.001). The bond strength between ceramic plates and resin cement in all test groups decreased after thermocycling (P < 0.001). All specimen in test group 1 and 2 lost bond, and the bond strength of test group 3 and 5 [(0.59 ± 0.17), (0.89 ± 0.84) MPa] were significantly lower than that of test group 4 and 6 [(14.63 ± 3.03), (16.64 ± 1.90) MPa], and the bond strength of test group 6 were significanlty higher than that of test group 4.</p><p><b>CONCLUSIONS</b>Sandblasting improves durability of bond between zirconia ceramic and resin cement containing MDP, not only by increasing the roughness and area of ceramic surface, but also by changing its surface element composition to obtain more chemical bond.</p>
Subject(s)
Aluminum Oxide , Chemistry , Ceramics , Chemistry , Dental Bonding , Dental Stress Analysis , Materials Testing , Microscopy, Electron, Scanning , Resin Cements , Chemistry , Shear Strength , Surface Properties , Zirconium , Chemistry摘要
<p><b>OBJECTIVE</b>To explore the synergetic effect of norcantharidin (NCTD) and adriamycin (ADR) on the proliferation and apoptosis of multiple myeloma (MM) cells.</p><p><b>METHODS</b>Human MM cell line U266 cells were treated with NCTD alone (10 µmol/L) or in combination with ADR (0.25 µmol/L). MTT and Annexin V/PI staining were used to determine cell viability and apoptosis. The protein expression of nuclear factor-κB P65 (NF-κB P65), phosphorylated NF-κB p65 (p-NF-κB p65), NF-κB P65 inhibitor IκBα, phosphorylated IκBα (p-IκBα), survivin, Bcl-2 and Bax were determined by Western blot. Immunohistochemistry was used to determine the expression of vascular endothelial growth factor (VEGF).</p><p><b>RESULTS</b>(1) NCTD potentiated the cytotoxicity and pro-apoptotic effects induced by ADR. The combination of NCTD and ADR had synergistic anti-proliferation effect. (2) Combination of ADR and NCTD downregulated the expression of nuclear NF-κB P65 and cytoplasm p-IκBα induced by ADR. The expression of nuclear NF-κB P65 and cytoplasm p-IκBα decreased from 2.08 ± 0.29 and 0.39 ± 0.07 to 0.48 ± 0.08 and 0.02 ± 0.01 respectively, while the expression of the cytoplasm NF-κB P65 and IκBα were unchanged in the ADR alone group and the combined group. (3) The expression of survivin and bcl-2 decreased from 0.31 ± 0.05 and 0.23 ± 0.05 to 0.03 ± 0.02 and 0.05 ± 0.02, while the expression of Bax increased from 0.46 ± 0.06 to 0.62 ± 0.08 respectively in ADR alone group and combined group. (4) The positive rate of VEGF in ADR group and combination group were (44.6 ± 4.4)% and (27.0 ± 2.1)% respectively, indicating that NCTD could potentiate the inhibition effect on VEGF induced by ADR.</p><p><b>CONCLUSIONS</b>The results suggest that NCTD can potentialize the chemosensitivity of multiple myeloma cells to ADR through regulating NF-κB/IκBα signaling pathway and NF-κB-regulated gene products including survivin, Bcl-2, Bax and VEGF.</p>