Objective To determine the effect of Leptospira interrogans lipopolysaccharide (L-LPS) inducing apoptosis of murine mononuclear-macrophage cell line( J774A. 1 ), and apoptotic regulation of Toll-like receptor(TLR) and associated intracellular signaling pathways. Methods Lipopolysaccharide (L-LPS) of L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai 56601 was prepared using phenol-water method. The effects of L-LPS inducing J774A. 1 cell apoptosis and the apoptosis-blocking with FasL neutralizing antibody were detected by flow cytometry. Real-time fluorescent quantitative RT-PCR (qPCR) and flow cytometry were performed to measure the changes of Fas/FasL mRNA and protein expression levels in J774A. 1 cells before and after L-LPS treatment. The regulations in L-LPS-induced cell apoptosis by TLR2 and TLR4 as well as p38MAPK, JNK, ERK pathways were determined by either TLR2 or TLR4 antibody blocking test, signaling pathway blocking test and flow cytometry. Results 56.50%, 69.28% and 24.35% of the J774A. 1 cells after treatment with 100 ng/ml L-LPS for4, 12 and 24 h were apoptotic,while the apoptosis rates were decreased to 11.21%, 21.58% and 12.70% after the cells blocked by FasL neutralizing antibody(P ＜0.05). The levels of FasL and Fas mRNAs in J774A. 1 cells treated with L-LPS for 4, 12 and 24 h were elevated with 1.34, 2.12, 2.10 times and 2.45, 3.87, 3.12 times compared to those in the L-LPS untreated cells (P ＜ 0. 05 ), respectively, while the expression rates of FasL and Fas proteins were upregulated to 18.61%, 60.13%, 42.75% and 76.34%, 85.70%, 77.92% from 4.82% and 15.32% apoptotic rates in the L-LPS untreated cells, respectively( P ＜0.05 ). The L-LPS-induced apoptosis rate( 11.54% ) of TLR2 antibody blocked J774A. 1 cells was significantly lower than that(66.56% ) of the J774A. 1 cells without TLR2 antibody blocking( P ＜0.05 ), but L-LPS-induced apoptosis rate of TLR4 antibody blocked J774A. 1 cells was as high as 55.27% ( P ＞ 0.05 ). Compared to the apoptosis rate (62.17%) in the p38MAPK and JNK pathway-free J774A. 1 cells, the L-LPS-induced apoptosis rates in p38MAPK blocked cells(20.54% ) and JNK blocked cells(47.98% ) were significantly lower( P ＜0.05 ),and the apoptosis rate in ERK blocked cells was as high as 61.72% ( P ＞ 0.05 ). Conclusion L-LPS was recognized by TLR2 and upregulates both Fas and FasL expression via p38MAPK and JNK pathways, which involving in the process of the L-LPS-induced cell apoptosis.

Objective To explore the expression of nuclear factor-erythroid 2-related factor 2(Nrf2) and the molecular chaperone of cytoplasmic Keap1 in premature newborn rats exposed to hyperoxia.Methods Completely randomized design method was performed,one-day old preterm SD rats were randomly divided into two groups:hyperoxia group and air group.The preterm SD rats in hyperoxia group were continuously exposed to oxygen(oxygen >0.85)and air group in room air.After 1 ,4,7,10,14 days of exposure,the pre-term SD rats of two groups were sacrificed,whole lung of these rats were isolated,the lung histological chan-ges were observed by HE staining.Total lung RNA was extracted,Nrf2 and Keap1 mRNA were detected by RT-PCR.Western-blot was used to detect the changes of Nrf2 protein expression.Results (1 )Compaired with air group,the expression of Nrf2 in lung tissue of hyperoxia group significantly increased after 4,7 days of exposure(4 d:0.314 ±0.064 vs.0.521 ±0.086,7 d:0.440 ±0.121 vs.0.658 ±0.076)(P <0.05 ),the general tendency decreased after 10 days,but the expression of Nrf2 became significantly weak after 14 days of exposure in hyperoxia group(P <0.05).(2)The expression of Keap1 mRNA in hyperoxia group signifi-cantly increased in 1 ,4 days(1 d:0.352 ±0.052 vs.0.547 ±0.075,4 d:0.363 ±0.074 vs.0.658 ±0.076) (P <0.05),the general tendency decreased after 7 days of exposure,but the expression of Keap1 mRNA in hyperoxia group became significantly weaker than its expression after 10,14 days of air group(P <0.05 ). (3)In comparison with air group,Nrf2 protein expression in hyperoxia group increased after 1 ,4 days of ex-posure with no significant difference(P >0.05),but had a tendency of decreasing after 7 days.On day 10, 14,its expression in hyperoxia group became significantly weak compared with that of air group(10 d:1.325 ±0.464 vs.0.755 ±0.348,14 d:1.662 ±0.474 vs.0.867 ±0.1 15 )(P <0.05 ).Conclusion Oxidation outbreak results in the abnormal expression of Nrf2 and Keap1 in the lung of premature SD rats induced by hyperoxia exposure,which adjusts the levels of oxidative stress in the body,these changes participate in the development of hyperoxia induced lung injury,the activity of Nrf2 may be increased by hyperoxia exposure, and alleviate hyperoxia lung injury in premature rats through antioxidation of Nrf2.

ObjectiveTo investigate the protective effect of ozone oxidative preconditioning (OzoneOP) on apoptosis induced by acute renal ischemia/reperfusion (I/R) injury in rats.Methods The right kidneys of rats in control group were excised.A rat I/R model was established in I/R group.In OzoneOP group,the renal OzoneOP was induced by rectal insufflation of 5.0～5.5 ml oxygen plus ozone (ozone 50 mg/L,1 mg· kg- 1 · d- 1,once every day).The parameters (blood urea nitrogen and creatine )of renal function were determined by auto-biochemical analyzer. The concentrations of malondialdehyde (MDA) and superoxide dismutase (SOD) were measured by using chromometry.Cytochrome c (CytC) was examined by using immunohistochemistry and Western blotting.The mRNA expression levels of TNF-α,IL-1β and IL-6 in the renal tissue were detected by using RT-PCR.ResultsCompared with the control group,the blood urea nitrogen,creatine,MDA and the release of CytC were increased significantly in I/R group.After OzoneOP,the levels of blood urea nitrogen,creatine,MDA,CytC and the mRNA expression levels of TNF-α,IL-1-β and IL-6were significantly decreased and SOD levels were significantly increased.ConclusionOzoneOP could relieve renal I/R injury of rats by improving antioxidation capability,cutting down cytokines contents and inhibiting the release of CytC from mitochondria.

Objective To observe the effects of Triptolide on the expression of toll-like receptor 4 (TLR4) in renal ischemia/reperfusion (I/R) injury in rats. Methods A renal I/R model was established. Rats were randomly separated into the following experimental groups. Group 1, shamoperated control (n = 15) : rats were subjected to surgical manipulation, without the induction of renal ischemia. Group 2, I/R (n = 18): rats were subjected to left renal ischemia for 45 min followed by reperfusion. Group 3, TRI + I/R (n = 18): Before the I/R procedure (as in group 2), rats were intraperitoneally injected with TRI (0.4 mg/kg), once every day, three times. Rats were killed at the 1st, 3rd, and 5th day after I/R injury. The parameters of renal function were determined by autobiochemical analyzer. The expression of TLR4 was detected by RT-PCR and Western blotting. Results As comparedwith the sham-operated control group, serum BUN and Cr levels were significantly increased in the rats undergoing I/R procedure at the 1st, 3rd, and 5th day (P＜0. 01). After the treatment with TRI, the levels of BUN and Cr and the expression of TLR4 in the renal tissues were significantly decreased (P＜0. 05). Conclusion TRI could relieve renal I/R injury in rats by inhibiting the TLR4 expression.

Objective To evaluate the precision,accuracy,linear range and anti-interference performance of HbA1c assay by u-sing Primus Trinity Ultra2 automated analyzer.Methods According to Document EP published in 2004 by American Clinical and Laboratory Standards Institution(CLSI)and relative literatures,the precision,accuracy,linear range and anti-interference perform-ance were evaluated.Results The within-run and between-run CVs of HbA1c determination were both less than that announced by manufacture.In the tests of samples with certain concentrations,all the results were within the verification range respectively.In the linear verification,regression equation between the theoretical and measured values demonstrated that r2 >0.95,and a value was within the range of 0.97-1.03.For Primus Trinity Ultra2 the limit of interference effect didn′t exceed the allowable error at the medical decision concentrations.Conclusion The precision,accuracy,linear range and anti-interference was evaluated of the Primus Trinity Ultra2 automating HbA1c all could meet the manufacture′s declaim and the clinical needs.

OBJECTIVE: To analyze the relationship between traditional Chinese medical syndromes and fungal pneumonia for moderate and advanced lung cancer patients. METHODS: We retrospected 115 moderate and advanced lung cancer patients with different syndromes in traditional Chinese medicine (qi deficiency, yin deficiency, blood deficiency, yang deficiency, blood stasis, phlegm dampness, phlegm heat, damp heat, cold dampness, qi stagnation, heat toxin), who had the concurrent fungal pneumonia, and used regression analysis method to analyze the data. RESULTS: When the patients had the phlegm heat syndrome, they got a significantly higher risk of having fungal pneumonia (P < 0.01); and when they had the heat toxin syndrome, they also had a high risk of having fungal pneumonia (P < 0.05). CONCLUSION: The phlegm heat and heat toxin syndromes are the risk factors for moderate and advanced lung cancer patients having concurrent fungal pneumonia.

Objective To evaluate the predictive value of urinary neutrophil gelatinase-associat-ed lipocalin (NGAL) and interleukin-18 (IL-18) for delayed graft function (DGF) in kidney transplan-tation. Methods Serial urine samples collected at 0, 12 and 24 h after operation from 86 kidney transplantation patients were analyzed by enzyme-linked immunosorbent assay for NGAL, IL-18 and RBP. Results Fifteen patients developed DGF. At 12 h after transplantation, the level of urine NGAL elevated significantly (1712.75±474.6 vs. 863.1±199.8 without DGF, P<0. 001). The in-creases of urine IL-18 (29. 2±4.1 vs. 28.7±4.2 without DGF, P>0. 05) was not significant. At 24 h, both urine NGAL(2905.0±1108.1 vs. 911.8±221.0 without DGF,P<0. 001) and IL-18(211.3± 34.0 vs. 86.9±22.8 without DGF, P<0. 001) increased significantly, whereas the changes of urine RBP and serum creatinine (SCr) were not significant. ROC analysis showed that the area under the curve of NGAL and IL-18 were 0. 90 and 0.76 respectively, the cut-off values were 996.5 ng/mg and 148.5 ng/mg, the diagnostic sensitivities in DGF were 90.2% and 76.3%, specificities were 82.6% and 66.4% respectively. Conclusions Both urine NGAL and IL-18 could potentially be early predic-tive marker of DGF. The level of NGAL elevated earlier than IL-18, which may be more effective in predicting DGF.

Objective·To investigate the correlation of single nucleotide polymorphism (SNP) of complement factor H (CFH) gene with schizophrenia in Chinese Han population.Methods·The genotype,allele,and haplotype frequencies of 5 SNP loci (rs800292,rs 1061170,rs 10801555,rs 10922096 and rs2019727) in CFH gene were compared between 418 patients with schizophrenia (case group) and 655 normal people (control group) by SNaPshot technique.Results·All SNP loci were well genotyped in the subjects.Correlation analysis showed that rs1061170 locus allele frequency distribution difference between case group and control group was statistically significant (corrected P=0.045),while genotype and allele frequencies of other SNP loci were not significantly different (all corrected P＞0.05).The frequency of haplotype C-A-T-A-A (rs800292-rs1061170-rs10801555-rs10922096-rs2019727) in case group was different from that in control group (corrected P=0.013).Conclusion·The allele polymorphisms of rsl061170 and the haplotype C-A-T-A-A of rs800292-rs 1061170-rs 10801555-rs 10922096-rs2019727 may be associated with schizophrenia in Chinese Han population.

Objective To investigate the interference of hyperlipidemia and hyperbilirubinaemia to HbA1c measurements by ion‐exchange high‐performance liquid chromatography(IE‐HPLC) method .Methods Fresh whole‐blood samples collected with EDTA‐K2 anticoagulant tubes were divided into four groups :control group(HbA1c<6 .2% ) ,diabetes group(HbA1c≥6 .2% ) ,hyperlipi‐demia group(TG 3 -20 mmol/L);hyperbilirubinaemis group (TBIL 21 -549 μmol/L) .HbA1c of these samples were measured with affinity chromatography(AC‐HPLC) and IE‐HPLC respectively .Results When HbA1c≤18 .7% ,r=0 .993 ;95% confidence interval(CI) of HbA1c results by using IE‐HPLC method was -0 .71 -0 .89 ;coefficient of variation was -5 .8% -6 .8% ;P=0 .198 and the difference was not statistically significant .When HbA1c< 16 .3% ,r= 0 .997;95% CI of HbA1c results with IE‐HPLC method is -0 .31-0 .67;coefficient of variation was -5 .8% -4 .3% .P=0 .000 and the difference was statistically signifi‐cant .No interference was detectded with the results ;When HbA1c was 16 .3% -18 .7% ,positive bias was observed with the re‐sults .When TG≤20 .78 mmol/L ,r=0 .995;95% CI of HbA1c results with IE‐HPLC method was -0 .26-0 .50 ;coefficient of var‐iation was -5 .5% -5 .8% .P=0 .000 and the difference was statistically significant .No interference was detectded with the re‐sults;When TBIL≤549 .3 μmol/L ,r=0 .990 ;95% CI of HbA1c results with IE‐HPLC method was -0 .08 -0 .63;coefficient of variation was -14% -4 .1% .P=0 .000 and the difference was statistically significant .When TBIL≤342 .1 μmol/L ,r= 0 .994 ;95% CI of HbA1c results with IE‐HPLC method was -0 .09-0 .50;coefficient of variation was -5 .5% -4 .1% .No interference was detectded with the results .When TBIL was 380 .7-549 .3 μmol/L ,negative bias was observed with the results .Conclusion Our data indicated that HbA1c measurement with IE‐HPLC method could resist the interference of hyperlipidemia;When TBIL≤380 .7 μmol/L and HbA1c<16 .3% ,the results could meet the needs of general clinical detection .Clinical staff should choose more specific HbA1c measurement method according to the patient's condition .

Objective? To explore the low frequency pulsed electrical stimulation on improving limbs pain among patients with diabetic peripheral neuropathy so as to provide an effective clinical basis for treatment and nursing of those patients. Methods? From March 2017 to March 2018, we selected 75 diabetic peripheral neuropathy patients with limbs pain in Chinese Surgery, the Shangqiu First People＇s Hospital as subjects by convenience sampling. All of the patients were randomly divided into control group (n=38) and experimental group (n=37) with the random number table. Patients of control group received routine treatment and nursing. On the basis of routine treatment and nursing, patients of experimental group were given the low frequency pulsed electrical stimulation. Finally, we compared the pain condition and effects of pain therapy of patients between two groups. Results? Before intervention, there was no statistical difference in the pain of patients between two groups (P＞0.05). After intervention, the limbs pain of patients in experimental group was lower than that in control group with a statistical difference (P＜0.05);the effects of limbs pain of patients in experimental group was better than that in control group with a statistical difference (P＜ 0.05). Conclusions? Low frequency pulsed electrical stimulation in Sanyinjiao and Yongquan point can regulate arteries, veins and Qi-blood, dredge the channels and collateral to make Qi-blood unobstructed so as to play a role in relieving the limbs pain of diabetic peripheral neuropathy patients. It is worth to be widely used in clinical application.