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Objective To explore the clinical efficacy and safety of low-intensity pulse ultrasound in the treatment of medial painful knee osteoarthritis.Methods A total of 10 patients with medial painful knee osteoar-thritis were collected and treated with low-intensity pulsed ultrasound.VAS visual analogue scores,WOMAC and Lysholm knee osteoarthritis rehabilitation scale scores were performed before treatment,immediately after treatment,1 week,2 weeks and 4 weeks after treatment.The efficacy and safety were compared before and after treatment.The ultrasound images before and after treatment were compared to verify its safety.Result VAS,WOMAC and Lysholm scores were statistically significant before and immediately after treatment,1 week and 2 weeks after treat-ment(P<0.05),but there was no statistically significant difference between them and 4 weeks after treatment(P>0.05).VAS decreased significantly before treatment,immediately after treatment,1 week after treatment and 2 weeks after treatment,and were(5.26±1.12),(2.76±0.92),(2.74±1.26),(3.42±0.93),respectively.The secondary outcome indicators of WOMAC and Lysholm were significantly changed before treatment,immediately after treatment,1 week after treatment and 2 weeks after treatment.They were(24.03±4.43)and(66.93±14.14),(12.75±1.63)and(79.86±10.82),(12.76±1.82)and(81.93±7.67),(15.17±2.73)and(77.08±9.73),respectively.No significant abnormality was found in the ultrasound images before and after treatment.Conclusion low-intensity pulsed ultrasound safely and effectively regulates the peripheral knee nerve in the treatment of painful knee osteoarthritis,providing a new idea for the intervention and treatment of clinical knee osteoarthritis.
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Objective Study of the rare hepatitis B virus patients model cases which both the hepatitis B surface antigen (HBsAg) and hepatitis B surface antibody (HBsAb) were positive, and discussion of its cause and the clinical value. Methods serum markers of hepatitis B virus (HBV-M) was detected by microparticle enzyme immunoassay chemiluminescence (MEIA); HBV-DNA was detected by fluores--cence quantitative PCR method, alanine aminotransferase and aspartate aminotransferase were detected by colorimetric, and all the data were combined with the clinical features of patients for comprehensive analysis. Results 1) HBsAg and HBsAb double positive detection rate was 2.3% in 15600 cases of hepatitis B patients, there was no significant difference in the positive rate of different sex groups and different age groups (P> 0.05); 2) HBsAg, HBsAb, HBeAb, HBcAb positive mode accounted for the highest proportion in all HBsAg and HBsAb double positive cases, the percentage was 57.9%; 3) the positive rate of HBV DNA in hepatitis B patients with HBeAg positive rate were higher than HBeAg negative group in all HBsAg and HBsAb double positive cases; and the incident rate of double variation nt 1762 A-T/nt 1764 G-A in HBeAg negative group was higher than that in HBeAg positive group. There were significant differences between two groups (P < 0.05); 4) the detection rate of HBsAg and HBsAb double positive in patients with chronic hepatitis B were higher than those of asymptomatic carriers, liver cirrhosis, hepatitis B and hepatitis B hepatocellular carcinoma (P < 0.05). Conclusion The phenomenon of both positive HBsAg and HBsAb does not indicate the elimination of the hepatitis B virus infection, but it is likely suggested the mutation of the virus. It is necessary to prompt clinical detection of serum HBV DNA, so as to determine whether the virus in patients is in the replication status, and it also provide some help for clinical individualized treatment of HBsAg and HBsAb double positive patients.
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Objective To investigate the effects of interferon alpha?2b(IFNα?2b)on serum Hepcidin in hepatitis C patients and its mechanism. Methods Hepatitis C patients were divided evenly into treatment group and control group according to whether they had received treatment with IFNα?2b in the past 3 months. The serum hepci?din was compared between the two groups. HepG2 cells and LO2 cells were treated for 24 hours at varied levels of IFNα?2b(0,50,100,200,400μL)and real?time PCR was used to detect the hepcidin,interleukin?6(IL?6)and signal transduction and transcription activator 3(STAT3)mRNA expression of cells. The protein levels of STAT3 and phosphorylated STAT3(pSTAT3)were measured by Western blot. The changes of these indexes were observed with the gradual increase of IFNα?2b levels. Results Serum Hepcidin level in the treatment group was significantly lower than the control(P<0.05). IFNα?2b inhibited the Hepcidin mRNA in HepG2 cells and LO2 cells. pSTAT3 was significantly decreased with the increased levels of IFNα?2b(P<0.05),and the expression of IL?6 and STAT3 had no significant changes with the increase of IFNα?2b. Conclusion The serum Hepcidin levels can be decreased because IFNα?2b suppresses the expression of Hepcidin,and its mechanism may be related with inhibited STAT3 pathway activation.
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Objective To investigate the role and the mechanism of ppk1 gene (coding for polyphosphate kinase 1) in oxidative stress resistance in uropathogenic Escherichia coli (UPEC).MethodsMutant strains with ppk1-deletion (△pk1) and complemented strains (△pk1-C) were constructed based on the UPEC strain CFT073.A comparative analysis was conducted to analyze survival rates of CFT073, △pk1 and △pk1-C strains at different time points while they were under oxidative stress.Differences in protein expression between CFT073 and △pk1 strains were analyzed using mass spectrometric analysis.Differences between CFT073 and △pk1 strains in expression of katG and katE genes were analyzed using real-time quantitative RT-PCR.Results The survival rate of △pk1 strains was lower than that of CFT073 strains at every time point, while the survival rate of △pk1-C strains was basically the same as that of CFT073 strains.Gel image analysis and mass spectrometric analysis revealed that six proteins were down-regulated and one was up-regulated in △pk1 strains as compared with those in CFT073 strains.Expression of the catalase-coding genes katG and katE in △pk1 strains were respectively (20.5±8.2)% and (20.9±6.9)% of those in CFT073 strains (P<0.05).Conclusion The ppk1 gene plays an important role in oxidative stress resistance in UPEC by modulating the expression of catalase-coding genes katG and katE.
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Objective To evaluate the diagnostic value of hepatitis C virus core antigen(HCV-cAg),hepatitis C virus(HCV-IgG) and hepatitis C virus(HCV-RNA) in the laboratory diagnosis of Hepatitis C.Methods HCV-cAg and HCV-IgG were detected by enzyme-linked immunosorbent assay(ELISA),HCV-RNA was detected by real-time fluorescent quantitative polymerase chain reaction(RT-PCR) in 84 suspected HCV patients and 87 healthy control subjects.Results In 84 suspected HCV patients,the HCV-IgG positive rate was 84.5%,HCV-cAg positive rate was 13.1%,HCV-RNA positive rate was 52.4%.Among 71 cases of HCV-IgG positive patients,there were 35 cases with negative HCV-RNA,the false positive rate was 49.3%.In 11 cases of HCV-cAg positive patients,there were 5 cases with negative HCV-RNA,the false positive rate was 45.5%.In 44 cases of HCV-RNA positive diagnosis of hepatitis C patients,HCV-IgG false negative rate was 18.2%,HCV-cAg false negative rate was 86.4%.The false negative rate of combined detection of HCV-cAg and HCV-IgG was 13.6%,and the true positive rate was 100.0%.Conclusion HCV-cAg and HCV-IgG have certain false negative and false positive in laboratory diagnosis of HCV,combine these two methods,or joint with HCV-RNA detection,could reduce the rate of missed diagnosis.
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Objective Using chemically synthesized small interfering RNA (siRNA) transfected HepG2.2.15 cells to construct a cell model in interfering hepatitis B virus (HBV) X gene, studying the inhibi-tion of HBV replication and antigen expression in vitro. Methods After transfection of HepG2.2.15 cell for 24 h, 48 h, 72 h, detecting the cell supernatant of HBsAg and HBeAg by chemiluminescence immunoassay, the cell supernatant HBxAg protein by ELISA , the HBx mRNA relative expression of transfected cell was detected by fluorescence quantitative polymerase chain reaction (PCR), the ability of cell proliferation was detected by CCK-8 assay. Results After HBx-siRNA transfected HepG2.2.15 cells, cell proliferation ability was inhibited. The cell of HBx mRNA and the cell supernatant of HBxAg expression decreased (P < 0.05); at the same time it in-hibited the expression of HBsAg and HBeAg. The suppressed peak and the inhibition rate were 66% and 58%respectively at 72 h. The fluorescence quantitative PCR confirmed that expression of HBV DNA in the super-natant was decreased. Conclusion The HepG2.2.15 cell interference model of HBV X gene has been success-fully constructed, which has an effect of inhibiting proliferation of HepG2.2.15 cells and replication and expres-sion of HBV gene in vitro.
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Objective To explore the role of ppk1 gene in E.coli CFT073 strain during urinary tract infection (UTI). Methods C57BL/6 mouse models of acute UTI with the wild-type(CFT073) and ppk1 mutant (△pk1) infected, were used to compare the bacteria colonization and inflammation induction abilities of bladder tissues. Results In the mouse models, the △pk1 strain showed a significantly lower infection rate (73.3% vs 93.3%) and lower adhesion frequency of bladder (0.01%vs 0.5%) than those of the CFT073 strain. The expression of IL-6 and TNF-αwere reduced in the bladder of △pk1 infected group (P<0.05). Hematoxylin-Eosin tissue staining showed that the damage degree of bladders in △pk1 infected mice were less serious than the CFT073 infected mice. Conclusion ppk1 gene plays an important role in E.coli colonization to bladder and the inflammation induction ability.
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Objective To investigate the effect of down-regulating of T cell immunoglobulin domain and mucin domain protein-3 (Tim-3) gene in asthmatic mouse model by short hairpin RNA(shRNA) and explore the role of Tim-3 on the differentiation of helper T lymphocytes 17 (Thl7),airway inflammation,as well as airway hyperresponsiveness in pathogenesis of asthma.Methods An asthmatic murine model was established by way of ovalbumin (OVA) sensitization and challenge.Treating Tim-3 gene was intranasally administered with Tim-3-specific shRNA.Invasive pulmonary impedance method was adopted to detect mice airway resistance.Flow cytometry analysis was performed to determine the levels of Th17 ; enzyme linked immunosorbent assay was performed to determine the concentrations of IL-17 and transforming growth factor-beta(TGF-β) in supematant.Results Asthmatic mice model was successfully established.By using Tim-3 shRNA silencing Tim-3,airway inflammation was reduced and airway hyperresponsiveness was declined in asthmatic group ;the levels of Th17 cells in asthmatic group [(6.43 ± 1.01)%] were significantly increased compared with normal controls[(1.75 ± 0.02) %].After using Tim-3 shRNA silencing Tim-3,the levels of Th17 cells [(2.36 ± 0.28) %] were decreased (F =40.05,P < 0.05) ; the level of IL-17 in asthmatic group [(118.8 ± 16.5) ng/L] was significantly increased compared with normal controls[(72.5 ± 13.6)ng/L],after using Tim-3 shRNA silencing Tim-3,the level of IL-17 [(73.6 ± 12.5) ng/L] was decreased (F =32.80,P < 0.05),while the expression of TGF-β did not change.Conclusions Down regulation of Tim-3 gene can decrease airway inflammation and airway hyperresponsiveness,which may be related to the change of Th17 cell differentiation.
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Objective To construct a Polyphosphate kinase 1 ( ppk1) gene deletion mutant of uro-pathogenic Escherichia coli (E.coli) CFT073, and to explore the biological properties of the mutant strain . Methods The ppk1 gene deletion strain (△pk1) was constructed based on CFT073 E.coli strain by usingλRed homologous recombination technology .A comparison analysis was conducted on adhesive and invasive abilities between CFT073 wild type strain and △pk1 strain in in vitro model of human bladder cancer epithe-lial cell 5637 .Crystal violet staining method was used to evaluate the influences of ppk1 gene deletion on biofilm formation.Results The CFT073 ppk1 deletion mutant strain was successfully constructed .Com-pared with the wild type strain ,△pk1 strain showed impaired adhesive and invasive abilities to 5637 cells. Moreover , the absorbance values of crystal violet at 570 nm at each time point of the mutant strain group were also lower than those of the wild-type strain group .Conclusion The ppk1 gene deletion mutant of uro-pathogenic E.coli CFT073 could be successfully constructed by Red homologous recombination technology and its biological properties indicates that ppk1 gene plays an important role in the pathogenesis of uropatho-genic E.coli infection through regulating the abilities of adhesion , invasion and biofilm formation .
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Objective:To develop the representative fingerprint for the quality control of placenta polypeptide injection.Methods:The chromatographic separation was performed using a Phenomenex Gemini C18 column (250 mm × 4.6 mm,5 μm) maintained at 30 ℃.0.1% aqueous trifluoroacetic acid (Solvent A) and acetonitrile contained 0.1% TFA (Solvent B) were used as mobile phase with a gradient elution.Detection wavelength was 280 nm with the sample injection volume of 50 μL; the flow rate was 1.0 mL/min.The fingerprints of different samples were investigated by similarity analysis.Results:Nine peaks were identified as the characteristic common peaks.The similarities of the fingerprints of the 10 batches of samples were above 0.992.Conclusion:This method showed high precision and good repeatability,and provided the basis for the improvement of the quality control of placenta polypeptide iniection.
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Morphological examination of blood cells is an important part of the hematology examination course. In order to enrich teaching resources, network of bilingual education resource was established and put into application by the Department of Hematology in Guangzhou Medical College. The repository improved teaching quality of cell morphology, and played a role in training personnel of hematology examination with solid basic skills.
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Objective To detect the distribution of H.pylori ureA, vacA s1 gene and cagA subtype(ABC, ABD, ABAB, AAD, et al) in patients with digestive diseases in Guangzhou and investigate the relationship with the pathological findings of gastric mucosa.Methods A total of 227 randomly selected gastric mucosa from patients with digestive diseases were enrolled in the research, including 46 without pathological changes, 130 with chronic gastritis, 29 with peptic ulcer, 15 with atrophic gastritis and 7 with gastric cancer.Real-time PCR assay were used to detect Helicobacter pylori ureA gene and vacA s1 gene.EPIYA motifs in the 3′ region of cagA were amplified by conventional PCR followed by subtype sequencing. The conserved gene ureA was used to detect H.pylori infection.Results Among the 227 patients with digestive diseases, 50.7% (115/227) patients were H.pylori positive, in which 91.3%(105/115) carried vacA s1 and 78.3% (90/115) carried cagA. Four types of cagA-EPIYA subtype were detected, including ABC 17.8%(16/90), ABD 78.9%(71/90), AAD 2.2%(2/90) and ABAB 1.1%(1/90).In the non-pathological change group, 32.6% (15/46) were H.pylori positive, in which 28.3% (13/46) carried vacA s1 and 26.1% (12/46) carried cagA;in chronic gastritis group, it was 48.5% (63/130), 43.8% (57/130) and 36.2% (47/130), respectively;in ulcer group, it was 72.4% (21/29), 65.5% (19/29) and 55.2% (16/29), respectively;in atrophic gastritis group, it was 66.7% (10/15), 66.7% (10/15) and 66.7% (10/15), respectively;in gastric cancer group, it was 85.7% (6/7), 85.7% (6/7) and 71.4% (5/7), respectively.The distribution of H.pylori among the 4 groups had statistical significance (χ2=16.72;P<0.01). H.pylori was more prevalent in ulcer, atrophic gastritis and cancer group than in inflammation group and non-pathological change group (χ2=16.02;P<0.01).In patients infected by H.pylori, there was no significant difference in the distribution of vacA s1 gene as high virulence factors among non-pathological change, inflammation, ulcer, atrophic gastritis and cancer group (χ2=2.00;P=0.74), as well as cagA (χ2=3.44;P=0.49) and EPIYA subtypes (χ2=3.66;P=0.45).Conclusions H.pylori infection is significantly associated with the pathological change of gastric mucosa for patients with digestive diseases in Guangzhou, while the relationship with the pathogenicity of H.pylori with high virulence genotype is not significant.More samples and diseases reclassification are needed to make an advanced analysis of the effect of H.pylori with high virulence in gastrointestinal diseases development.
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0 05). Between the patients with high level of serum HBV-DNA and the ones with low level of serum HBV-DNA, both the quantity and positive rate of HBV-DNA in PBMC had a significant difference (P