ncing analysis validated that all four-type base-quenched probes could provide unbiased genotyping results ( Kappa =1, P=0.00), although. Conclusion This method is simple, economic and suitable for large-scale genotyping studies.

AIM:To determine whether the vitreous cavity (VC) is capable of supporting the induction of deviant immune response to retinal soluble (S) antigen and to observe the influence of interleukin-1 (IL-1) on the immunologic properties of the VC. METHODS: Retinal S antigen was inoculated into the anterior chamber (AC) and VC in Wistar rats. Seven days after antigen inoculation, the recipient animals were immunized with S antigen and complete Freund's adjuvant. Delayed-type hypersensitivity (DTH) was then assessed by footpad challenge. To alter systemic immune conditions, IL-1 was administrated by intraperitoneal injection. RESULTS: Antigen-specific DTH did not develop in rats in which S antigen was injected into the AC and the VC. In contrast, strong DTH was elicited by S antigen injected into the AC and VC if IL-1 was administrated systemically for 7 consecutive days after the antigen challenge. CONCLUSION: The VC is capable of supporting immune deviation to soluble antigen by actively suppressing antigen-specific DTH. Systemic administration of exogenous IL-1 eliminates the capacity of the VC to support immune deviation inducing by soluble antigen injected locally.

Objective Experimental autoimmune anterior uveitis (EAAU) is a useful model for human anterior uveitis. The purpose of this study was to observe whether EAAU development could be prevented by anterior chamber-associated immune deviation (ACAID). Methods Bovine melanin protein (BMP) in phosphate-buffered saline (PBS) was injected into the anterior chamber of the right eye of rats; control animals were injected with PBS alone. Seven days later, all animals were immunized with BMP in complete Freund's adjuvant (CFA) and pertussis toxin. The severity and incidence of uveitis was monitored by clinical examination and histopathology. The delayed-type hypersensitivity (DTH) responses were evaluated by footpad swelling elicited by injection of BMP.Results Rats intraocularly injected with BMP showed a reduced severity and incidence of EAAU, and significantly suppressed DTH response compared to control rats.Conclusion These data suggest that the ACAID procedure can be utilized to prevent the development of EAAU.

Objective To study the association of biallelic polymorphism at position 196 in exon 6 of tumor necrosis factor receptor 2(TNFR2) gene and coronary artery disease(CAD).Methods A retrospective case control study was designed in this investigation.Patients with angiographically proven CAD (n=142)and coronary atherosclerosis(n=215)were compared to healthy volunteers(n=631). Genotyping was performed by ShineRoar probes analysis.Results People with GG genotype at position 196 in exon 6 of TNFR2 gene have more risks of development of CAD(OR=2.556,95% CI:1.051～6.218, χ2=4.57,P=0.033)and CA(OR=2.547,95% CI:1.162～5.579,χ2=5.81,P=0.016) than those with TT genotype.Further analysis after adjustment for sex and age demonstrated that GG genotype was not associated with CAD(OR=0.614,95% CI:0.166～2.279,χ2=0.53,P=0.466)and coronary atherosclerosis(OR=O.644,95%CI:0.200-2.069,χ2=0.55,P=0.459).Conclusion Tumor necrosis factor receptor 2 gene polymorphism at position 196 in exon 6 was not an independent risk factor of development of CAD and coronary atherosclerosis among Han Chinese.

In this study,a synthesized quadruple antigenic epitope gene region of the classical swine fever virus (CSFV)E2 glycoprotein was expressed in E.coli to a obtain target protein.This target protein was used as a coating antigen to establish an indirect ELISA for specifically detecting anti-CSFV antibodies in serum samples from pigs.The P/N cut-off value of this assay was 1.92 by receiver operating characteristic curve(ROC)analysis based on 30 negative sera and 80 positive samples.The test gave 97.5% sensitivity and 96.7% specificity compared with the indirect hemagglutination(IHA)test.The inter-assay and intra-assay coefficients of variation (CVs)for 16 sera were both ≤6.8%.No cross-reactivity between the coating antigen and anti-bovine viral diarrhoea virus(BVDV)antibodies was observed.

Objective To detect the expression level of human zinc finger 23 (ZNF23) in hepatocellular carcinoma tissue samples and HepG2 cell lines and investigate the relationship between hZNF23 expression and clinicopathological characteristics of HCC and cell apoptosis. Methods The expression levels of hZNF23 and GAPDH mRNA in 37 cases of HCC were measured by real-time RT-PCR. The association between the expression of hZNF23 and the clinicopathological characteristics of HCC was analyzed. Cultured HepG2 cells were divided into 4 groups ( control group, 1.25 μg/ml , 2.5 μg/ml and 5 μg/ml cisplatin)or 6 groups( control group, 1.25 μg/ml, 2.5 μg/ml, 5 μg/ml, 10 μg/ml and 20 μg/ml cisplatin). MTT method was employed to evaluate cell proliferation. Annexin V-FITC assay was used to assess percentage of apoptotic HepG2 cells. The expression levels of hZNF23 and GAPDH mRNA of HepG2 cells after apoptosis induced by cisplatin with a series of concentrations were measured by real-time RT-PCR.Results The median ( quartile1, quartile 3) expression levels of hZNF23 mRNA in 37 HCC tissue samples and adjacent tissue samples were 8.84 (3.59-15.05), 22.20 ( 13.85-42.90 ), respectively. There was significant difference ( U = 259.5, P ＜ 0.01 ). The median ( quartile1, quartile 3 ) expression levels of hZNF23 mRNA in cancer tissue samples with Edmondson stage Ⅰ + Ⅱ [12.80(4.80-19.50)] was much higher than those in stage Ⅲ + Ⅳ [5.01 ( 2.88-11.68 ), U = 99.00, P ＜ 0.05] The median ( quartile1,quartile 3 ) expression levels of hZNF23 mRNA in patients with and without hepatic cirrhosis were 9.92(3.80-15.25) , 3.21 (2.78-3.60), respectively. The median ( quartile1, quartile 3 ) expression levels of hZNF23 mRNA in HBV infection and non-infection patients was 9.09(3.72-15.25 ), 2.48 (1.79-12.10),respectively. There was no significant difference between groups with and without hepatic cirrhosis and between HBV infection and non-infection groups( U = 16. 00 and 24.00, P ＞0.05 ). MTT assay indicated that cisplatin significantly inhibited HepG2 cells proliferation in a dose-dependent manner. Annexin V-FITC/PI assay showed that HepG2 cells apoptosis rates were (0.9 ± 0.2 ) %, ( 4. 2 ± 0.3 ) %, ( 9.8 ± 4. 3 ) %,(23.0 ± 6.0)%, respectively. Cisplatin significantly induced HepG2 cells apoptosis in a dose-dependent manner( F = 27.89, P ＜ 0.01 ). The expression levels of hZNF23 mRNA in cisplatin groups [( 10.39 ±3.08) × 10-5, (24.10 ± 2.09) × 10-5, (6.90 ± 2.24) × 10-4] were significantly lower than that of the controlgroup[(94.80±1.80) ×10-5, F=6.027, P＜0.01]. Conclusions The expression level hZNF23 mRNA is related to Edmondson stage of HCC. The apoptosis effect of cisplatin on HepG2 cells may be associated with the upregulation of hZNF23.

Objective To investigate the clinical application of multiplex allele-specific PCR assays for simultaneous detection of the mitochondrial 12S rRNA A1555G and C1494T mutations associated with aminoglycoside-induced hearing impairment.Methods Three standard plasmids of different genotypes (wild-type, A1555G mutant and C1494T mutant) were constructed for templates and allele-specific primers aiming directly at wild-type and mutant of mitochondrial DNA nt1555 and nt1494 were designed for developing a multiplex allele-specific PCR technique to detect the A1555G and C1494T mutations.Then the method was applied to clinical screening of 138 non-syndromic hearing loss subjects and confirmed by DNA sequencing.Results Multiplex allele-specific PCR was successfully applied to the detection of A1555G and C1494T mutations in a cohort of 138 Han Chinese genetically unrelated hearing-loss subjects.Finally, 11(7.97%) unrelated affected subjects harbored the A1555G and C1494T mutations in the 12S rRNA gene(10 cases for A1555G and 1 cases for C1494T), which was well consistent with results of DNA sequencing [7.97%(11/138), Kappa=1.000, P＜0.01].Conclusion This study indicates that the multiplex allele-specific PCR assay is useful, convenient and reliable in the detection of the A1555G and C1494T mutations, which could identify the subjects at risk and effectively prevent of aminoglycoside-induced hearing loss.

OBJECTIVETo prepare zirconia toughened nano-composite alumina ceramic powder for dental application. Physical and chemical property of the prepared material were tested, and the effect of development technology on composite powder was also studied in this study.

METHODSNano-composite alumina powder was prepared by surface-induced precipitation method. The effect of pH value and dispersing agent content on volume of alumina suspension sediment was recorded. The effect of ultrasonic time on agglomeration was measured also. X ray diffraction (XRD) was used to analyze powder phase before and after the stabilizer was added. Scanning electronic microscope (SEM) was applied for characterizing the specimen.

RESULTSThe dispersion was better at pH=9 and wt (dispersing agent) = 0.2% approximately 0.3%. Selecting proper ultrasonic time can decrease the agglomeration of powders and lower the average particle size. XRD analysis indicated that the phase composition of the prepared nano-composite ceramic powder was shown as alpha-Al2O3, t-ZrO2 and a small amount of m-ZrO2 after the addition of stabilizer. Through SEM observation, nanometer-sized ZrO2 particles (80 approximately 100 nm) were uniformly located on the surface of submicrometer alumina grains.

CONCLUSIONSBy choosing appropriate preparation method, weakly agglomerated powders with fine particle size can be obtained. The zirconia part of nano-composite powder was transmitted to partially stabled zirconia after the use of stabilizer.

OBJECTIVETo compare the analgesic effect between multimodal and patient-controlled intravenous analgesia(PCIA) in patients with rheumatoid arthritis(RA) in the perioperative period of knee joint replacement.

METHODSFrom June 2015 to June 2016, 40 RA patients undergoing total knee arthroplasty were randomly divided into two groups. There were 20 patients in PCIA group, including 3 males and 17 females, with an average age of(59.6±2.3) years old, who received controlled instillation of sufentanil analgesia controlled by an intravenous analgesia pump. There were 20 patients in multiple model analgesia group, including 2 males and 18 females, with an average age of(56.3±1.3) years old, who were treated with continuous femoral nerve block, local injection of knee joint and combined buprenorphine patches. The VAS score and the incidence of adverse reactions and HSS score were compared between the two groups after operation. The advantages and disadvantages of the two modes of analgesia were evaluated.

RESULTSOn the 6 th and 24 th hours after surgery, the VAS scores of the multimodal analgesia group were significantly lower than those of the PCIA group(<0.01). On the 48 th hour after surgery, the VAS scores was significantly lower in the multimodal analgesia group than those in PCIA group(<0.000 1), both in the state of motion and at rest. On the 1 st week after surgery, the HSS score of the multimodal analgesia group was significantly higher than that in the PCIA group(<0.000 1). The pain score and the degree of activity in HSS score of the multimodal analgesia group were better than those in PCIA group (<0.05). The functional score of multimodal analgesia group was significantly better than that of PCIA group(<0.01). But there was no significant difference in muscle strength scores between two groups.

CONCLUSIONSMultimodal analgesia is an ideal analgesic plan for total knee arthroplasty TKA patients with RA in perioperative period, which has good effects and little adverse reaction.

OBJECTIVETo investigate the morphological characteristics of the mandibular first premolars in people from Pearl River Delta region in Guangdong province using three techniques, including periapical radiographs, the radiographs with files inserted the canals and the clearing technique.

METHODSA total of 363 extracted mandibular first premolars were collected and numbered. Two preoperative radiographs were taken in buccolingual and mesiodistal directions respectively. After access opening, the files were placed in the canals and two other radiographs were taken. The mandibular first premolars with multi-canal system were selected and observed under dental operating microscope (DOM). The mandibular first premolars were made transparent and were categorized using the Vertucci's classification.

RESULTSThere were different results among the three approaches. Periapical radiographs could be used to distinguish only between one and multiple canals systems. The incidence of multiple canals was 33.33% from the radiographs with file. The mandibular first premolars had a high frequency (34.44%) of multi-canal system by clearing method. The root canal morphology of the mandibular first premolars showed great variance. The canal orifices of the mandibular first premolars with one or two canal distributed in a buccolingual line. The floor of pulp chamber of the mandibular first premolars with three or four canals was a plat form.

CONCLUSIONThe mandibular first premolars have a high frequency multi-canal system and could be classified in many categories. Using DOM and radiographs with file is a useful way in judging the canal numbers and categories.