Objective To explore the potential value of multiple tumor markers chip( C- 12) in preoperative diagnosis of gastric cancer. Methods The serum levels of 12 rumor markers were measured in 45 gastric cancer patients, 38 benign gastrosis patients and 65 normal controls by use of C-12 in order to find out the most levels of CA199, CEA, CA242, AFP and CA125 in the gastric cancer patients were significantly higher than those of the benign gastrosis patients and normal controls. Moreover, the serum levels of β- HCG and HGH were also significantly higher in gastric cancer group than benign gastric disease group and control group ( P ＜sis of gastric cancer. CEA is the TM with the highest sensitivity, validity and negative predictive value of 57.8% ,81.8% and 77.1% ,respectively whereas CA242 is the TM with the highest specificity and positive CEA + CA125 + CA199 and CEA + CA242 + CA199 + CA125, respectively. The sensitivity, specificity and validity of the best combination of 2 TMs, 3 TMs and 4TMs for gastric cancer were not statistically significantly different from those of C-12 and the best TM ( P ＞ 0.05 ). Conclusion The multiple tumor markers chip ( C-12 ) has a relatively high value in the preoperative diagnosis of gastric cancer. The best combinations of 2 TMs ( CEA + CA125) ,3 TMs ( CEA + CA125 + CA199 ) and 4TMs ( CEA + CA242 + CA199 + CA125 ) for gastric cancer diagnosis could be sufficient to replace the combination of 12 TMs.

Objective To evaluate the role of transient receptor potential ankyrin 1 (TRPA1) in the dorsal root ganglion neurons in the development of diabetic neuropathic pain (DNP) in rats.Methods Twenty-four Sprague-Dawley rats with DNP were randomly divided into 3 groups (n-=8 each) using a random number table:DNP group,TRPA1-specific siRNA group (siRNA group) and TRPA1-negative siRNA group (NC group).Another 8 Sprague-Dawley rats with normal blood glucose served as control group (C group).In siRNA group,TRPA1-specific siRNA 45 μl was injected intrathecally.In NC group,TRPA1-negative siRNA 45 μl was injected intrathecally.In DNP and C groups,normal saline 45 μl was injected intrathecally.On 2nd day after intrathecal administration,the lumbar segment (L4-6) of the dorsal root ganglions was removed for determination of the expression of TRPA1 mRNA.On 7,14,21 and 28 days after intrathecal administration (T1-4),MWT was measured.Results Compared with DNP group,TRPA1 mRNA expression was down-regulated in siRNA and C groups.Compared with DNP group,and MWT was significantly decreased at T1.2 in siRNA group,MWT was decreased at T1-3 in NC group,MWT was increased at T1-4 in group C.Compared with siRNA group,MWT was significantly increased at T1-4 in group C.MWT was significantly higher at T1~ in group C than in NC group.Conclusion TRPA1 in the dorsal root ganglion neurons is involved in the development of DNP in rats.

Objective To evaluate the clinical value of multiple tumor marker protein chip in diagno-sis and detection of postoperative recurrence of breast cancer.Methods The serum levels of 12 tumor makers (CA199,NSE,CEA, CA2A2,Ferritin,β-HCG,AFP,f-PSA,PSA,CA125,CA153 and HGH)were measured in 70 preoperative breast cancer patients, 32 recurrence patients,52 non-recurrence patients and 76 normal con-trois.Results ①The breast cancer group had significantly higher positive rate than that of the controls (P<0.05).The positive rates and serum levels of CA199,CEA,CA242,Ferritin,CAI25 and CA153 in breast cancer patients had those of control significant differences compared with groups (P<0.05).②The recurrence group had significantly higher positive rate than that of non-recurrence group (P<0.05).The positive rates and se-rum levels of CA199, CEA, Ferritin, CA125 and CA153 in the recurrence patients had significant differences compared with those of non-recurrence patients(P<0.05).③The positive rate of recurrence group had signif-icant difference compared with that of breast cancer group(P<0.05).Moreover,The positive rate and serum level of Ferritin in the recurrence patients had significant difference compared with that of breast cancer pa-tients.Conclusion The multiple tumor marker protein chip detective system has valid value of clinical appli-cation in the diagnosis and detection of postoperative recurrence of breast cancer.The combination detection of CA199, CEA, Ferritin ,CA125 and CA153 may be the economical and effective in the diagnosis and detection of postoperative recurrence of breast cancer.

The effect of recombinant human phospholipase D2(rhPLD2)in vivo was investigated on the secretion of serum glycosyl phosphatidylinositol-specific phospholipase D(GPI-PLD)in guinea pigs of chronic asthma．Ater treating the guinea pigs attacked by chronic asthma with rhPLD2,the GPI-PLD activity detection was canrried out by phase separation of human placental alkaline phosphatase in Triton X-114．Compared with the healthy guinea pigs(NS group),the serum GPI-PLD in the guinea pigs of chronic asthma are much higher than that of control groups,P≤0．01．Our results showed that rhPLD2 could significantly reduce the secretion of GPI-PLD when the guinea pigs were attacked by chronic asthma．

Objective To evaluate the changes in the expression of acetylated histone H3 (Ac-H3) and deacetylase silent information regulator 1 (SIRT1) in dorsal root ganglions in a rat model of negative phenotype neuropathic pain.Methods Forty-eight male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 2 groups (n =24 each):sham operation group (group S) and C-fiber dysfunction group (group CFD).The rats were anesthetized with 10％ chloral hydrate 3 ml/kg.C-fiber dysfunction was induced by exposing sciatic nerve to 8％ capsaicin for 30 min in group CFD.The thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) were measured before and on 1,3,7 and 14 days after CFD.Six rats were then sacrificed at each time and the lumbar segments (L5) of the dorsal root ganglions were removed for detection of SIRT1 mRNA expression (by RT-PCR) and Ac-H3 and SIRT1 protein expression (by Western blot).Results Compared with group S,TWL was significantly increased at 1,3,7 and 14 days after CFD,SIRT1 mRNA and protein expression was up-regulated and Ac-H3 expression was down-regulated at 3,7 and 14 days after CFD (P ＜ 0.05),while no significant change was found in MWT at each time point in group CFD (P ＞ 0.05).Conclusion The mechanism of negative phenotype neuropathic pain is related to up-regulation of deacetylase SIRT1 expression and decreased acetylation of histone H3 in rat dorsal root ganglions.

Objective To clone and characterize Profilin encoding genes in Amaranthus spinosus and to analyze the contribution of different amino acids in isoallergens to allergen antigenicity and tertiary structure. Methods The primers were designed according to the core sequences which were obtained by bioinformatic analysis of the known Profilin amino acid sequences, followed by gene cloning from the Ama- ranthus spinosus cDNA pool and subsequent confirmation by double-digestion, colony PCR and DNA sequen- cing. Antigenicity evaluation and tertiary structural modeling of the encoded protein were accomplished by online software MULTIPRED and SWISS-MODEL, respectively. Results Two panallergenic genes, named as PRF7 and PRF23, were acquired from Amaranthus spinosus. Sequence and structure analysis demonstra- ted that there was some discrepancy in tertiary structures of the encoded proteins, besides distinct difference in their amino acid sequences. PRF7 exhibited high homology with panallergen Profilins Q64LH0, with the identities 98%, whereas the homology of PRF23 and Q9XF42 (apple allergen) was 81%. Q64LH0 and PRF23 were modeled as 3nulA (Q42449) and lg5uB (Q9LE18), respectively. PRF23 exhibited distinct0 three dimensional structural difference in certain fragments compared with Q64LH0 and other Profilins. Though the average values of antigenicity displayed no difference between Q64LH0 and PRF23 on whole se- quences, the antigenicity of PRF23 on certain fragments was obviously lower than that of Q64LHO because of the alteration of some amino acids with different characters, implying the cause of lower incidence of hay fe- ver in South China than in North China. Conclusion Based on sequence analysis, antigenicity evaluation and tertiary structural modeling for Q64LH0 and PRF23, we obtained lots of useful information about the contribution of different amino acids to antigenicity and protein structures, thus would facilitate allergen ge- netic improvement by amino acid replacement.

Objective To investigate the effects of repeated low dose intravenous infusion of low molecular weight iron dextran and iron sucrose on oxidative stress in chronic renal failure (CRF) rats. Methods CRF model was established by 5/6 subtotal nephrectomy (5/6 Nx). Four weeks after removing the right kidney, successful rats were randomly divided into low molecular weight iron dextran group, sucrose iron group and CRF control group. The sham group was established simultaneously. The dose of iron administrated in each rat was similar in iron dextran group and sucrose iron group. There were 6 rats in each group. Animals were observed for 6weeks, then the blood, urine and renal tissue samples were collected, and indexes of renal function,anemia, iron status and oxidative stress were investigated. Results The hemoglobulin (Hb) level in iron groups was significantly higher as compared to control group (P＜0.05) but was not significantly different between two iron groups. The levels of serum iron, ferritin and saturation rate of transferring (TS) were obviously lower in control group as compared to sham group (P＜0.05).Levels of above 3 indexes were significantly higher in two iron groups as compared to control group (P＜0.05), but were not significantly different between two iron groups. Concentration of plasma advanced oxidation protein products (AOPP) was obviously higher in two iron groups than that in control group [(127.84±21.19) μmol/L, (134.21±29.38) μmol/L vs (81.83±19.93) μmol/L, P＜0.05]. Plasma malonaldehyde (MDA) was significantly higher in iron sucrose group than that in iron dextran group [(6.06±0.73) nmol/L vs (4.99i0.80) nmol/L, P＜0.05]. Serum levels of superoxide dismutase (SOD) and total anti-oxidant capacity (TAOC) had no significant differences among three CRF groups. Concentration of plasma glutathione peroxidase (GSH-Px) was significantly decreased in three CRF groups as compared to sham group (P＜0.05), while plasma GSH-Px was significantly lower in sucrose iron group than that in iron dextran group and control group [(2123.11±74.78)nmol ·ml-1 ·min-1 vs (2352.84±163.90) nmol· ml-1 ·min-1, (2310.23±125.99) nmol ·ml-1 ·min-1, P＜0.05]. Conclusions Injection of intravenous iron can partially improve the anemia and the iron status indexes in 5/6 Nx CRF rats. Repeated low dose intravenous infusion of iron dextran and iron sucrose can aggravate the oxidative stress state in CRF rats, and the iron sucrose is worst.

Objective To compare the efficacy of the conventional PPD skin test and a new enzymelinked immunospot assay(TSPOT-TB)for diagnosing latent tuberculosis infection(LTBI)in patients with rheumatic diseases.Methods Two hundred and sixty rheumatic patients were enrolled,and all were screened for LTBI based on clinical history,chest X-ray,PPD skin test or TSPOT.Results The positive rate of TSPOT assay was 24.1%and that of PPD skin test was 39.4%.The overall concordance rate between the 2tests was 61.0%.Among PPD negative patients (n=149).29 were TSPOT(+)(19.5%).Among PPD(+)patients(n=98),69 were TSPOT(-)(70.0%).The patients who got BCG vaccination or had history of tuberculosis infection showed a significantly higher rate of positive result of PPD skin test than those who did not (P<0.05 or P<0.01).While in TSPOT assay,the BCG vaccination or history of tuberculosis infection did not show influence on TSPOT results(P>0.05).Of the 127 patients who received biological agents after screening for LTBI,9 patients were pretreated with isoniazide.Twenty-seven patients stopped biological agent treatment because of the positive results of PPD or TSPOT.Twenty three patients who had positive PPD but negative TSPOT results received biological agent treatment without isoniazide,and none of them developed active tubereulosis after 6 to 18 months of follow-up.Conclusion BCG vaccination affects the result of PPD test in rheumatic patients,but has no influence on TSPOT results.The infection rate of latent tuberculosis of rheumatic patients in our research is 23.8%detected by TSPOT.

The association of urinary cytokine concentration with the incidence of urinary tract infection in patients with diabetes mellitus was explored. Urinary interleakin-6 (IL-6) level in 156 female type 2 diabetes mellitus patients was tested and followed up for 6 months. The incidence of urinary tract infection between the top quartile of baseline urinary cytokine concentration with the bottom quartile was compared. The incidence of top quartile of urinary IL-6 concentration at baseline was significantly lower than that of bottom quartile ( 13.5% vs 37.8%, P＜0.05 ). Abnormality of urinary IL-6 concentration may be involved in the mechanism leading to higher prevalence of urinary tract infection in patients with diabetes mellitus.

Objective To investigate effect of the venom peptide liquor on adjunctive arthritis (AA) in mice .Methods The male Kunming mice were randomly divided into 6 groups ,named as the control group ,model group ,positive control group(1 mg/kg dex-amethason) ,wine group(10 mL/kg) ,low dosage of venom peptide liquor group(8 .3 mL/kg) and high dosage of venom peptide liq-uor group(33 .2 mL/kg);the right toes ,ankle diameter and whole body weight were measured at 7-day intervals ;the spleen index , serum level of circulating immune complexes(CIC) were determined after the thirty-fourth day when the mice was put to death .Re-sults The level of CIC in AA mice decreased significantly (P<0 .05) ,the decrease of spleen index and the reduction of toe and an-kle swelling in the low dose group were significant(P<0 .01) .Conclusion The venom peptide liquor exhibited apparent inhibitory effect on AA in mice .