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Objective:To explore the effect of Bushen Huatan prescription on serum lipopolysaccharide （LPS） and Toll-like receptor 4 （TLR4）/ myeloid cell differentiation protein 88 （MyD88）/nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） signaling pathway in rats with ovariectomy-induced osteoporosis. Method:Sixty SPF 6-month-old female rats were randomly divided into sham operation group， model group， estradiol valerate group and Bushen Huatan prescription low， medium and high dose groups.One week after modeling by bilateral ovariectomy， 8 rats in each group were selected to receive intragastric administration.The estradiol valerate group was given 0.184 mg·kg<sup>-1</sup> by gavage， and Bushen Huatan prescription low， middle and high dose groups were given 4.7， 9.4 and 18.8 g·kg<sup>-1</sup> by gavage， sham operation group and model group were given 0.9% saline 4 mL by gavage respectively.After 12 weeks of intervention， the rats were sacrificed for detection.Serum LPS was detected by enzyme linked immunosorbent assay （ELISA）， while protein expressions of TLR4， MyD88 and phosphorylated （p）-NF-<italic>κ</italic>B p65 in bone tissue were detected by Western blot， and the mRNA expressions of TLR4， MyD88， NF-<italic>κ</italic>B p65， IL-1<italic>β</italic>， and IL-6 in bone tissue were detected by quantitative real-time polymerase chain reaction（PCR）. Result:Compared with sham operation group， the serum LPS level as well as protein expression of TLR4， MyD88， p-NF-<italic>κ</italic>B p65 and mRNA expression of TLR4， MyD88， NF-<italic>κ</italic>B p65， IL-1<italic>β</italic>， and IL-6 significantly increased in model group（<italic>P</italic><0.05）.Compared with the model group， serum LPS level， protein expression of TLR4， MyD88， and p-NF-<italic>κ</italic>B p65， mRNA levels of TLR4， MyD88， and NF-<italic>κ</italic>B p65 in bone tissues as well as downstream inflammatory factors IL-1<italic>β</italic>， IL-6 mRNA expression decreased to different degrees in estradiol valerate group and Bushen Huatan prescription high dose group（<italic>P</italic><0.05）. Conclusion:Bushen Huatan prescription can reduce serum LPS content， regulate mRNA and protein expression of TLR4， MyD88， NF-<italic>κ</italic>B p65 and p-NF-<italic>κ</italic>B p65 in TLR4/MyD88/NF-<italic>κ</italic>B pathway， and down-regulate mRNA levels of IL-1<italic>β</italic> and IL-6 in bone tissues to improve bone microstructure and inhibit the development of postmenopausal osteoporosis （PMOP）.
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Objective:To study the effect of Bushen Huatan prescription on helper T cell 17 （Th17）/T regulatory cells （Treg） balance of immune T cell subsets in the prevention and treatment of postmenopausal osteoporosis. Method:Sixty 6-month-old female SD rats were randomly divided into sham operation group， model group， estradiol valerate group （0.184 mg·kg<sup>-1</sup>） and Bushen Huatan prescription low， medium and high groups （4.7， 9.4， 18.8 g·kg<sup>-1</sup>） according to the random number table. All the groups except the sham operation group received ovariectomy to make postmenopausal osteoporosis model. Intragastric administration was started 1 week after operation， and the rats in model group and sham operation group received equal volume of normal saline， once a day for 12 weeks. Microcomputed tomography （Micro CT） was then used to detect bone mass and microstructure of rats， the contents of Forkhead box protein （Foxp3） and retinoic acid related nuclear orphan receptor （ROR<italic>γ</italic>t） in serum were detected by enzyme-linked immunosorbent assay （ELISA）， the mRNA expression levels of Foxp3 and ROR<italic>γ</italic>t in bone tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot was used to detect the protein expression of Foxp3 and ROR<italic>γ</italic>t in bone tissues， the number of Th17 and Treg cells in each group was analyzed and compared by flow cytometry. Result:Compared with the sham operation group， the bone mass and trabeculae of the model group decreased （<italic>P</italic><0.01）， the bone microstructure was destroyed， the concentration of Foxp3 in serum decreased， the concentration of ROR<italic>γ</italic>t increased （<italic>P</italic><0.01）， the mRNA and protein expression levels of Foxp3 in bone tissues decreased， ROR<italic>γ</italic>t increased， the number of Treg cells in bone tissues decreased， number of Th17 cells increased （<italic>P</italic><0.01）， and Th17/Treg ratio increased （<italic>P</italic><0.01） in model group. Compared with the model group， the bone mass in each treatment group increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， Foxp3 concentration in serum increased， ROR<italic>γ</italic>t concentration decreased （<italic>P</italic><0.01）， the mRNA and protein expression levels of Foxp3 in bone tissues increased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）， but no statistical difference was shown in mRNA expression between low dose group and the model group. In addition， the mRNA and protein expression of ROR<italic>γ</italic>t decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， number of Treg cells increased， number of Th17 cells decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， and Th17/Treg ratio decreased in treatment groups （<italic>P</italic><0.01）. Conclusion:Bushen Huatan prescription can increase bone mass， improve bone microstructure， increase the number of Treg cells and decrease the number of Th17 cells in ovariectomized rats. It is concluded that Bushen Huatan prescription may play a role in preventing and treating postmenopausal osteoporosis by regulating Th17/Treg balance.
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Objective To study the expression of CD269 and CD317 antigens in bone marrow cells of patients with multiple myeloma (MM),analyze its correlation with the laboratory indexes reflecting the progression of MM and evaluate its value in clinical diagnosis and treatment.Methods 63 newly diagnosed MM patients were selected as the study group by a casecontrol study.The expression rate of CD269 and CD317 in bone marrow blood of 35 patients with iron deficiency anemia and other antigens in bone marrow blood of 63 patients with MM were detected by flow cytometry.The levels of serum hemoglo bin (Hb),serumβ2-MG(β2-MG) and lactatedehydrogenase (LDH) in patients with MM were dctectcd,and the levels of CD269 and CD317 were analyzed statistically.Results The positive rates of CD269 in the study group and control group were (86.6±2.35)％ vs (4.33±l.69)％,rcspectivcly (t =4.256,P＜0.05)).The positive rate of CD317 was (71.42+ 0.62)％ vs (8.32+ 3.89)％,the difference was statistically significant (t=3.102,P＜0.05).In other expression,the expression level of CD269 and CD317 in CD56 positive group was significantly higher than that of negative group (t=4.032,P＜0.05),while the expression of CD117 the level of positive group was significantly lower than that of the negative group (t 2.832,P＜0.05),CD19,CD20 expression was not statistically significant difference between the two groups (P＞ 0.05).The levels of CD269 and CD317 in patients with MM were positively correlated with the level of CD56 expression (r =0.392,P＜0.05),and negatively correlated with the level of CD117 expression (r=-0.210,P＜0.05).The levels of CD269 and CD317 in patients with MM were significantly lower than those in the negative group (t=3.012,P＜0.05) and the levels of serum LDH in the positive group were lower than those in the negative group (t=2.024,P＜0.05).There was a negative correlation between Hb content (r=-0.212,P＜0.05) and negatively correlated with serum β2-MG (r=-0.312,P＜0.05).Conclusion The high expression of CD269 and CD317 in bone marrow cells in MM patients is related to the increase of CD56 and decrease of CD117 in patients with MM.
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Objective To explore the differential expression of CD269 and CD317 in patients with multiple myeloma(MM). Methods Newly diagnosed samles from patients of MM(20 cases)and iron deficiency anemia(20 cases),40 cases in total (from 06/2015 to 08/2013,the Department of Hematology,Central Hospital of Zhuzhou City)were collected.Real-time quantitative PCR(RQ-PCR)tests were used to detect the relative expression of CD269 and CD317 in bone marrow sam-ples,and the results were statistically analyzed with clinical features.Results The relative expression levels of CD269 and CD317 in patients with multiple myeloma(4.418±4.568,4.327±2.876)were significantly higher than those in the control group(0.600±0.838,1.033±1.335),the difference was statistically significant(t=3.676,4.646,all P<0.05)respective-ly,while not related with the gender,age(P>0.05).There was no correlation between the expression of CD269 and CD317 (r=0.041,P=0.864),but positively correlated with the ratio of myeloma cells(r=0.495,P=0.026;r=0.533,P=0.016).Conclusion CD269 and CD317 were highly expressed in patients with multiple myeloma and may be involved in the pathogenesis of multiple myeloma.
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Objective To determine the methylation level of P15INK4B gene promoter in different types of myelodysplastic syndromes (MDS) and its correlation with its prognosis.Methods Methylation frequency of the P15INK4B gene promoter in 44 cases of MDS were determined by methylation-specific polymerase chain reaction (PCR) and pyrosequencing,and its correlation with clinical classification and characteristics of MDS were statistically analyzed.Results Frequency of P15INK4B gene promoter methylation in myelodysplastic syndromes-refractory anemia with excess blasts Ⅱ (MDS-RAEB Ⅱ) patients was (46.89 ± 15.41) ％,significandy higher than that in other types of MDS (P ＜ 0.05),but no difference in promoter methylation frequency was detected among the other types of MDS (P ＞ 0.05) ; frequency of P15INK4B gene promoter methylation was found to be correlated with decline in platelet upon diagnosis (t =9.02,P ＜ 0.01),but showed no significant correlation with drop of hemoglobin or leukopenia (P ＞0.05).As for the correlation between P15INK4B gene promoter methylation and MDS risk stratification,no significant difference was detected between the low-risk and very low-risk groups (P ＞ 0.05),but significant differences were detected among the medium-risk,high-risk,and very high-risk groups (P ＜ 0.05).In addition,frequency of P15INK4B gene promoter methylation was (49.21 ± 8.78)％ in MDS patients that developed leukemia in the following two year,significantly higher than that in MDS patients who didn't (19.64 ± 6.24) ％ (P ＜ 0.05).Conclusions P15INK4B gene promoter methylation frequency is a valuable indicator of prognosis of MDS patients.
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Objective:To observe the effect of Pidotimod and Thalidomid to enhance the immune response and protective immunity induced by the epitope Vaccines From W2b2a of Toxoplasma gondii in mice.Methods:Mice were immunized intramuscularly with Pidotimod and pcDNA3-W2b2a,thalidomide and pcDNA3-W2b2a,respectively,then the immune response and the survival time of mouse attacked by Toxoplasma gondii were observed.Results:After the immunization,the level of IFN-γin sera of mice inculated with pcDNA3-W2b2a and Pidotimod,pcDNA3-W2b2a and Freund adjuvant,pcDNA3-W2b2a and Thalidomid were significantly higher than pcDNA3-W2b2a (all P<0.05).After the immunization,IgG,CD4+/CD8+T cell ratio ,proliferation of T cell induced by pcDNA3-W2b2a and Pidotimod,pcDNA3-W2b2a and Freund adjuvant were higher than pcDNA3-W2b2a,pcDNA3-W2b2a and thalidomid ( all P<0.05).After challenged with highly virulent tachyzoites,the mean survival time in immunized groups were significantly longer than control group (all P<0.05).Conclusion:Pidotimod ,Thalidomid adjuvant can increase protective immunity of epitope Vaccines From W2b2a of Toxoplasma gondii in mice.
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<p><b>OBJECTIVE</b>To study the relationship between hepatocellular carcinoma (HCC) and the interaction of polymorphisms in the NAD(P)H:quinone oxidoreductase (NQO1) gene with environmental factors using a hospital-based case-control study. FMETHODS: our-hundred newly diagnosed HCC cases and 400 healthy individuals (non-tumor controls) were enrolled in the study. Demographic information and medical history was obtained by questionnaire. TaqMan minor groove binder real-time PCR was carried out to detect the NQO1 C609T genotype using blood-derived DNA from all study participants. Unconditional logistic regression analysis was carried out to estimate the odds ratios (ORs) and 95% confidence intervals (CIs).</p><p><b>RESULTS</b>The frequencies of NQO1 609 CC, CT and TT genotypes were 23.75%, 50.25% and 28.00% in the HCC group, and 37.55%, 43.75% and 18.25% in the control group. The differences between the HCC and control group reached statistical significance (all P less than 0.05). The ORs of NQO1 609 CT and TT genotypes were significantly higher compared to the CC genotype; the adjusted OR(95% CI) of CT was 2.106(1.137-3.110) and of TT was 2.564(1.357-4.744). Individuals carrying the NQO1 609 T allelic gene had a significantly higher risk of HCC than those carrying the C allelic gene; the adjusted OR(95% CI) was 1.86(1.235-2.980). Interactions were found between hepatitis B virus infection with hepatitis B surface antigen (HBsAg)-positivity and NQO1 gene polymorphisms (adjusted OR: 2.431) and history of cancer (adjusted OR: 8.3592).</p><p><b>CONCLUSION</b>The NQO1 C609T genotype is associated with increased risk of HCC. Interactions between HBsAg-positive infection, history of cancer, and NQO1 gene polymorphisms may contribute to HCC.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Epidemiology , Genetics , Case-Control Studies , China , Epidemiology , Genotype , Liver Neoplasms , Epidemiology , Genetics , NAD(P)H Dehydrogenase (Quinone) , Genetics , Polymorphism, Single Nucleotide , Risk FactorsABSTRACT
<p><b>OBJECTIVE</b>To establish the methods of calculating and analyzing the multi-coefficient of variation significance test for the toxicology study.</p><p><b>METHODS</b>The paper aimed to confirm the significance level with the method of Bonferroni and then compared the methods of calculating and analyzing of the experiment groups with the control group respectively.</p><p><b>RESULTS</b>The significance level of multi-coefficient of variation significance test was confirmed as alpha1=0.0167. Compared with the control groups, the activity of ALT in serum both in 30 mg/kg and 60 mg/kg groups did not change in the average significance test, which was not statistically significant (P>0.05), while it changed in the variation significance test, which was of statistical significance (P<0.0167). The activity of AST in serum in 60 mg/kg group did not change in the average significance test (P>0.05), while it changed in the variation significance test (P<0.0167).</p><p><b>CONCLUSION</b>The complete changes of the indexes can only be shown by use of both the average significance test and the variation significance test together.</p>
Subject(s)
Animals , Female , Rats , Alanine Transaminase , Blood , Aspartate Aminotransferases , Blood , Disease Models, Animal , Lead Poisoning , Rats, Wistar , Statistical DistributionsABSTRACT
<p><b>OBJECTIVE</b>To study the correlation of eating raw fish with primary hepatic carcinoma (PHC), and to investigate the synergistic effect of HBV infection, alcohol consumption and eating raw fish on the oncogenesis of PHC.</p><p><b>METHODS</b>A hospital-based case-control study was conducted among 500 PHC patients and 500 non-cancerous patients in order to compare the history of eating raw fish. The synergistic pathogenetic action of eating raw fish, HBV infection and alcohol consumption on carcinogenesis of PHC was analyzed by crossover analysis and multiple logistic regression.</p><p><b>RESULTS</b>The rates of eating raw fish in the past between the case (54.8%) and the control group (8.4%) were significantly different (P < 0.001). OR value of suffering PHC in the patients who ate raw fish in the past was 13.6 (95% CI: 9.1-19.5) when compared with the non-cancerous patient. HBV infection, alcohol consumption and eating raw fish showed an interactive effect on the development of PHC, with a relative excessive risk of interaction(RERI) of 195.3 and 17.8; attributable proportion of interaction (API) of 0.8630 and 0.5251; and synergy index (S) of 7.5 and 2.8, respectively.</p><p><b>CONCLUSION</b>A history of eating raw fish may be an important risk factor for suffering primary hepatic carcinoma. HBV infection, alcohol consumption and eating raw fish may have a synergistic effect on the developing of primary hepatic carcinoma.</p>
Subject(s)
Adolescent , Adult , Aged , Animals , Female , Humans , Male , Middle Aged , Young Adult , Alcohol Drinking , Carcinoma, Hepatocellular , Epidemiology , Virology , Case-Control Studies , China , Epidemiology , Eating , Fishes , Hepatitis B , Liver Neoplasms , Epidemiology , Virology , Logistic Models , Odds Ratio , Risk Factors , SeafoodABSTRACT
<p><b>OBJECTIVE</b>To explore etiologic fraction (EF) and interaction of hepatitis B virus (HBV) infection and other risk factors for primary hepatocellular carcinoma (PHC) in Guangxi, China.</p><p><b>METHODS</b>A hospital-based case-control study including 500 PHC patients and 500 nontumorous patients was carried out in Guangxi. EF and interactions of HBV infection and other risk factors for PHC were analyzed by crossover analysis and nonconditional multiple logistic regression.</p><p><b>RESULTS</b>HBV infection, family history of PHC, diabetes mellitus, eating raw fish, heavy alcohol consumption, frequently used drug, low income, mental oppression and blood type B all were risk factors for PHC. With EFs of 0.725, 0.186, 0.119, 0.486, 0.385, 0.438, 0.277, 0.607, 0.299, respectively and with etiologic fractions attributable to interaction [EF(A xB)] of 0.736, 0.643, 0.849, 0.551, 0.592, 0.618, 0.902, 0.577; and indices of interaction of 0.743, 0.651, 0.853, 0.560, 0.600, 0.626, 0.907, 0.586, respectively.</p><p><b>CONCLUSION</b>Main risk factors for PHC might include HBV infection, family history of PHC, diabetes mellitus, eating raw fish, heavy alcohol consumption, frequently used drug, low income, mental oppression and blood type B. HBV infection with other risk factors might exert synergistic action on developing PHC and increase the risk of PHC.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , China , Epidemiology , Data Interpretation, Statistical , Liver Neoplasms , Risk FactorsABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between clonorchiasis and primary hepatocellular carcinoma (HCC) and analyze the synergistic actions of HBV infection, alcohol consumption and clonorchiasis on HCC development.</p><p><b>METHODS</b>This hospital-based case-control study was conducted among 444 HCC patients (cases) and 500 non tumor patients (controls) to compare the prevalence of clonorchiasis in the cases and the controls. The risk of clonorchiasis and the synergistic actions between HBV infection, alcohol consumption and clonorchiasis on HCC development were analyzed by crossover analysis and multiple logistic regression.</p><p><b>RESULTS</b>The prevalence of clonorchiasis in the cases (16.44%) was much higher than that of the controls (2.40%) (X2 = 56.58, P less than 0.01). In the case group, the OR value of those with clonorchiasis was 8.00 (95% CI: 4.34-14.92). The OR value was 4.82 (95% CI: 2.32-10.26) for the subjects whose clonorchiasis was diagnosed less than 10 years before their diagnosis of HCC, and was 17.54 (95% CI: 5.47-57.18) for those whose HCC was diagnosed more than 10 years ago. HBV infection, alcohol consumption and clonorchiasis showed an additive interaction in the development of HCC, with a relative excess risk of interaction of 110.43 and 18.23; attributable proportion of interaction of 0.80 and 0.63; synergy index of 5.18 and 2.84, respectively.</p><p><b>CONCLUSION</b>Clonorchiasis could be an important risk factor for HCC. When the course of clonorchiasis is prolonged, the risk of HCC could increase. HBV infection, alcohol consumption and clonorchiasis might have synergistic actions on the development of HCC.</p>