Objective To study the expression and clinicopathological significance of Paxillin and CAⅨ in the benign and malignant lesions of gallbladder.Methods The surgical resected specimens of 108 cases of gallbladder adenocarcinoma, 46 cases of peritumoral tissue, 15 cases of adenomatous polyp and 35 cases of chronic cholecystitis were routinely made paraffin embedded sections.The expressions of Paxillin and CAⅨ were stained with Envision immunohistochemistry.Results The positive rates of Paxillin and CAⅨ expression was significantly higher in gallbladder adenocarcinoma (60.2% and 49.1%) than those in peritumoral tissues (26.1%, x2 =15.00, P ＜0.01 and 23.9%,x2=8.41,P ＜0.01), adenomatous polyp (20.0%,x2=8.60,P＜0.01 and 20.0%,x2 =4.49,P＜0.05) and chronic cholecystitis(14.3% ,x2 =22.89, P＜0.01 and 11.4%,x2 =15.63,P ＜0.01).All the gallbladder epithelia of the benign cases with Paxillin and CA Ⅸ positive expression showed moderate to severe atypical hyperplasia.The positive expression rates of Paxillin and CA Ⅸ were significanctly lower in the cases of well-differentiated, maximal diameter of mass less than 2 cm, no lymph nodes metastasis and no surrounding tissues invasion than those of the cases with poorly differentiated, maximal diameter of mass over 2 cm, lymph nodes metastasis and surrounding tissues invasion.With Kaplan-Meier analysis, it suggested that the survival period after the surgery in Paxillin and CAⅨ positive expression cases was shorter than that of negative expression cases (x2 = 5.65,P＜0.05,5.65=5.92, P＜0.01).With multivariate Cox regression analysis, it indicated that both Paxillin and CAⅨ positive expression was an important indicator of the poor prognosis in gallbladder adenocarcinoma.Conclusion The expression of Paxillin and CA Ⅸ may be closely related to the carcinogenesis, tumor biological behaviors, and prognosis of gallbladder adenocarcinoma.The positive expression of Paxillin and/or CAⅨ is associated with poor prognosis.

ObjectiveTo study the expression of ABCG2, SFRP2, BRMS1 and HPA and detect their clinicopathologicalsignificancesinthebenignandmalignatntlesionsofthegallbladder.MethodsEnVisiom immunohistochemical method for determining the expressions of ABCG2, SFRP2,BRMS1 and HPA was used in paraffin-embedded sections of surgical resected specimens from gallbladder adenocarcinoma (n =108), peritumoral tissues (n =46 ), adenomatous polyp (n =15 ), and chronic cholecystitis ( n =35 ).ResultsThe positive rates of ABCG2 and HPA expression were significantly higher in gallbladder adenocarcinoma than that in peritumoral tissues, adenomatous polyp and chronic cholecystitis (P ＜ 0. 05 or P ＜ 0. 01 ) ; The positive rates of SFRP2 and BRMS1 expression were significantly lower in gallbladder adenocarcinoma than that in peritumoral tissues, adenomatous polyp and chronic cholecystitis(P ＜0. 05 or P ＜0. 01 ). The positive cases of ABCG2 and/or HPA as well as negative ones of SFRP2 and/or BRMS1in the benign lesions showed moderately-or severely-atypical hyperplasia of gallbladder epithelium. The frequency of samples with positive staining for ABCG2 and/or HPA in cases with small tumor volume (diameter ＜ 2 cm), no lymph node metastasis, and no invasion into surrounding tissues was significantly lower than that in cases with larger tumor volume (diameter＞ 2 cm ), lymph node metastasis, and invasion into surrounding tissues ( P ＜ 0.05 or P ＜ 0. 01 ). However, compared with ABCG2 and/or HPA, the expression of SFRP2 and/or BRMS1 showed an opposite correlation in these cases ( P ＜ 0. 05 or P ＜0. 01 ). Unitivariate Kaplan-Meier analysis showed that increased expressions of ABCG2 (P =0. 019) and HPA ( P =0. 016) or decreased expression of SFRP2 ( P =0. 019) and BRMS1 ( P =0. 008 )were associated with poorer overall survival, respectively. Multivariate Cox regression analysis showed that increased expression of ABCG2 (P =0. 018 ) and HPA ( P =0. 019) and/or decreased expression of SFRP2 (P =0. 015 ) and BRMS1 ( P =0. 011 ) were independently poor-prognostic predictors in gallbladder adenocarcinoma.ConclusionsThe expression of ABCG2, SFRP2, BRMS1 or HPA might be closely related to the carcinogenesis, clinical biological behaviors, and prognosis of gallbladder adenocarcinoma.

Objective To detection the methylation of p16INK4A in primary hepatocellular carcinoma, a nested bisulfite sequencing and methylation-specific polymerase chain reaction (BS-MSP) protocol was designed and used.Methods Bisulfite-modified DNA were amplified to evaluate the quality of templates with a pair of bisulfite sequencing primers in the first round of PCR, then subjected to methylation assay with corresponding methylation or unmethylation specific PCR primers.Representative PCR products were sequenced to confirm its correctness.Results 3 of 40 cases (7.5%) were failed to assay due to poor quality of templates, and 29 of 37 cases (78%) were detected p16INK4A methylation.Sequencing results confirmed that templates were correctly amplified.Conclusion BS-MSP technique might be valuable for methylation study on carcinogenesis and clinical assay.

Objective To investigate the effects of interleukin?17A on kidney injury induced by paraquat(PQ). Methods Seventy?two ICR mice were randomly divided into 3 groups:NS,PQ,and PQ+Ab (n=24 for each). The PQ?poisoning model was established by administering a gavage of PQ solution;mice in the PQ+Ab group were then administereda dose of anti?IL?17A antibody 2 hours later by i.p. injection,whereas the NS group were administered a corresponding volume of normal saline instead.The mice were killed at 8,24,48,or 72 h to obtain renal tissues and serum. An enzyme?linked immunosorbent assay(ELISA)was used to determine serum IL?17A,serum creatinine(SCr),and blood urea nitrogen (BUN)levels.Chemical colorimetry was used to detect the viability of myeloperoxidase(MPO )in renal tissue,and hematoxylin?eosin(HE)stain?ing was used to observe the renal pathologic changes. Immunohistochemistry(IHC)and PCR were used to examine IL?17A expression in renal tis?sues. Results Serum IL?17A,renal tissue MPO viabilities,BUN,and SCr were increased in the PQ and PQ+Ab groups,compared to those in the NS group(P<0.01). However,the above?mentioned parameters were lower in the PQ+Ab group than in the PQ group(P<0.01). Conclusion IL?17A promotes mouse kidney injury induced by acute PQ?intoxication through activating and/or recruiting neutrophils;therefore,blockade IL?17A,with antibody can attenuate the injury.

Objective To investigate the role of interleukin-17A (IL-17A) in acute paraquat (PQ)-induced lung injury in mice.Methods A total of 120 healthy SPF grade ICR male mice were randomly (random number) divided into three groups (n =40 in each):normal saline control group (NS),PQ poisoning group (PQ) and antibody neutralization group (PQ + Ab).Mice of PQ group and PQ + Ab group were given 5 mg/mL PQ by one gavage in a dosage of 25 mg/kg body weight,and 5 μg IL-17A neutralizing antibody intra-peritoneally administered immediately after PQ poisoning in PQ + Ab group;Equivalent volume of normal saline instead of PQ was given to mice of NS group.Six survival mice from each group were taken for experiment at 8 h,1 d,3 d,5 d,7 d after PQ poisoning:Wet to dry ratio (W/D) of lung was determined in mice of each group.HE staining of lung tissue was used to observe the histopathological changes under the light microscope and the pathological scores were graded;Serum interleukin-17A (IL-17A),interleukin-22 (IL-22),interleukin-6 (IL-6),transforming growth factor-β (TGF-β) were detected with enzyme linked immunosorbent assay (ELISA);Expression of interleukin-23 receptor (IL-23R) in lung tissue was determined with immunohistochemical;real-time fluorescence quantification PCR (qRT-PCR) was used to detect the expression of retinoic acid related solitary nuclear receptors' mRNA in lung tissue.Results After administration of PQ,W/D ratio increased (P ＜ 0.01),lung injury was observed in mice of PQ and PQ + Ab groups,levels of cytokines (IL-17A,IL-22,IL-6 and TGF-β) in serum elevated (P ＜0.05),and the expressions of IL-23R mRNA and RORγt mRNA increased (P＜0.01).But in PQ +Ab group,W/D ratio decreased (P ＜0.05),lung injury was alleviated,the levels of cytokines (IL-17A,IL-22,IL-6 and TGF-β) decreased (P ＜ 0.05),and the expressions of IL-23R mRNA and RORγt mRNA reduced (P ＜ 0.05).Conclusions Since IL-17A involves in the lung injury of the mice induced by acute paraquat poisoning,blockade of IL-17A significantly alleviates the acute lung injury in mice.

Drugs, Chinese Herbal ; Information Storage and Retrieval ; Medicine, Chinese Traditional

Objective To observe the changes of antithrombin activity(AT) and D-dimer in acute leukemia(AL)children complicated with disseminated intravascular coagulation(DIC) and to explore the changes of blood coagulation and fibrinolysis function.Methods Twenty-seven AL children without DIC were selected as AL group and 25 childern complicated with DIC were selected as observe group,36 health children were as control group.Plasma level of AT,D-dimer,fibrinogen,activated partial thromboplastin time and prothrombin time were tested by color substrate,immuno-latex turbidimetry,and coagulation method.And the rusults of AL group were compared with observe group and control group by SPSS 10.0 software.Results PT was significantly prolonged and the D-dimer in AL group and observation group were significantly higher than those in control group(Pa

[Objective]To explore the possible effects of the apolipoprotein A5(APOA5)gene rs2075291 and rs3135507 poly-morphisms on plasma lipid levels and the severity of coronary stenosis in patients with coronary heart disease(CHD)in Chinese Han people.[Methods]Polymerase chain reaction-restriction fragment length polymorphism method(PCR-RFLP)was used to identify the genotypes of the rs2075291 or rs3135507 polymorphism for the 324 patients with CHD and the 152 CHD-free controls,and the lipid levels between the genotypes were compared. The severity of coronary stenosis was assessed by the Gensini scoring system ,and the associations of the rs2075291 and rs3135507 polymorphisms as well as other factors with the Gensini scores were analyzed by mul-tivariate linear regression.[Results]The CHD patients had higher prevalence of hypertension ,and higher levels of triglycerides (TG),total cholesterol(TC),apolipoprotein B100(APOB100),lipoprotein(a)[Lp(a)],TG/high-density lipoprotein cholesterol (HDL-C),TC/HDL-C,low-density lipoprotein cholesterol(LDL-C)/HDL-C and APOB100/apolipoprotein AI(APOAI),and low-er levels of HDL-C and APOAI than the controls(P < 0.05 for all). The rs2075291 or rs3135507 allele and genotype frequencies were similar between the CHD group and the control group. The subjects with the rs2075291 GT genotype had higher levels of TG and TG/HDL-C,and lower levels of HDL-C than the subjects with the GG genotype in controls(P < 0.05 for all). In CHD patients, those with the rs2075291 GT genotype had higher levels of TG/HDL-C and TC/HDL-C than the patients with the GG genotype(P<0.05 for both). Regarding the rs3135507 polymorphism,the A allele carriers had lower level of APOB100 than the patients with the GG genotype(P<0.05). No significant association between the rs2075291 or rs3135507 polymorphism and the Gensini scores were detected by multivariate linear regression(P > 0.05).[Conclusion]The APOA5 rs2075291 polymorphism had significant effects on plasma lipid levels,but no effects on the occurrence and development of CHD. Further multicenter case-control studies with large samples are needed to validate these findings.

Objective To study on the effect of indicator animals in plague surveillance throngh detecting F1 antibody against Yersiniapestis(Y.pestis)in indicator animals in wild rat plague foci,provide scientific evidence for plague control and determining the range of epidemic area.Methods According to investigation scheme of wild rodents plagne foci in Yunnan Province,indicator animals Canis familiarils and Felis catu(C.familiarils and F.catus)related to the plague were investigated in 75 villages,14 township and 10 counties around Yulong County,and living rodents were captured by cage,sera of indicator animals and rodents relevant to plague were simultaneously collected and detected for F1 antibody against Y.pestis using indirect hemagglutination(IHA).Results Seropositivity rate of indicator animals were 6.76%(202/2987),being 24.69% in C.familiaris and 24.69% in F.catus,there were statistical significance(X2＝87.32,P＜0.01)between C familiaris and F catus,the latter beingmore than the former.But F1 antibody of rodents sera were not detected,its seropositivity rate was zero.there was a statistical significance(P＜0.01)between indicator animals and rodents.Conclusions Through serocpidemiological survey of indicator animals,new wild rat plague natural focus has been confirmed in YuLong County and Gucheng District in LiJiang City,therefore,serocpidemiological surveillance of indicator animals is very important for plague control and prevention.

Tripterygium glycosides preparation which extracted from the traditional Chinese herb Tripterygium wilfordii (TWHY), was widely used to treat the autoimmune diseases. Previous works demonstrated that TWHF had potent anti-inflammatory and immunosuppressive properties. But the different quality and high incident rate of side effects of different manufactures inhibited its clinical application. Since TWHF had been generally known to play a therapeutical effect by synergism of multiple constituents, it was necessary to build the relationship between the HPLC fingerprint and bioactivity so as to ensure the quality safety and efficacy. The HPLC fingerprint showed that description and content of peaks from different manufactures were diverse. Only 11 common peaks were found. In this study, mice spleen cells stimulated by Con A were used to test the proliferation inhibition bioactivity of TWHF preparations, which were incubated with 30, 15, 7.5, 3.75, 1.88 and 0.94 mg x L(-1) TWHF preparations for 48 h. The results showed that mice spleen cells proliferation was inhibited by all TWHF preparations significantly compared with the control group, which suggested the TWHF preparations showed immune suppress activity. The TWHF preparations from 7 manufacture showed different IC50 value, which might belong to different contents which showed in the HPLC fingerprint. Moreover, a relationship between the HPLC fingerprint and the bioactivity were established to identify important constituents by grey relational analysis (GRA). The result showed that all the contents were relative with the IC50, especially No. 5 and 10 peaks, but No. 1 peak, which was proved to be triptolide, had few contribute to the inhibition of mice spleen cells proliferation. The study of relationship between the HPLC fingerprint and the IC50 by GRA could help to investigate mechanism of bioactive and provide an evidence for the quantification of multi-constituents.
Animals ; Anti-Inflammatory Agents ; analysis ; pharmacology ; Cell Proliferation ; drug effects ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Glycosides ; analysis ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; drug effects ; Tripterygium ; chemistry