Objective: To investigate the influence of serum creatinine (sCr) at different time-points on prognosis of critically ill patients with acute kidney injury (AKI). Methods: This study was retrospectively analyzed the clinical data of critical patients with AKI who admitted to the mixed ICU of Xiaolan Hospital of Southern Medical University during March 2015 and January 2016. According to the clinical prognosis, the patients were divided into the renal replacement therapy (RRT) group and non-renal replacement therapy (non-RRT) group, 28-day renal loss group and renal recover group, hospital death group and survival group. Serum Cr at different time-points and clinical data were collected. The receiver operating characteristic (ROC) curve and the area under curve (AUC) were used to evaluate the capability of sCr at different time-points in predicting clinical prognosis. Results: During the study, 85 AKI patients were enrolled. The in-hospital mortality was 20%, RRT rate was 15.3%, and renal lose at 28 days after ICU admission was 31.8%. The levels of sCr out of ICU (o-sCr) and the peak of sCr were significantly higher in the RRT group than the non-RRT group (P<0.01). The o-sCr and p-sCr predicted RRT during the hospital stay with a higher AUC value (0.806 vs 0.833) than b-sCr and a-sCr. The levels of b-sCr, a-sCr, o-sCr, and p-sCr were all significantly higher in the28-day renal loss group than the kidney recover group (P<0.01). The levels of o-sCr and p-sCr were significantly higher in the hospital death group than the survival group (P<0.05). The o-sCr predicted renal lose at 28 days and hospital death with the highest AUC value of 0.850 and 0.797, respectively. Conclusions It cannot be ignored to monitor dynamically the level of sCr at different time-points in critical patients with AKI, which can predict the clinical prognosis such as hospital death, RRT and renal lose at 28 days after ICU admission.

Objective To investigate the correlation between TYMS gene mRNA expression level and EGFR mutation in stage Ⅲ A-N2 non-small-cell lung cancer tissue (NSCLC).Methods The branched DNA-liquidchip technology (bDNA-LCT) and mutant-enriched liquidchip (MEL) technology were used to detect the TYMS mRNA expression and EGFR mutations at exon 19 and 21 in NSCLC tissues from 30 patients with stages Ⅲ A-N2 NSCLC,and the results were analyzed.Results Among 30 cases,low TYMS mRNA expression was in 14 cases (46.7 ％),middle to low TYMS mRNA expression in 7 cases (23.3 ％),middle mRNA expression in 7 cases (23.3 ％),middle to high TYMS mRNA expression in 0 case and high TYMS mRNA expression in 2 cases (6.7％);12 cases of EGFR mutation were detected out with the mutation rate of 40.0％,including 6 cases of exon 19 deletion and 6 cases of exon 21 missense mutation.The TYMS mRNA expression level was correlated with the EGFR mutation.The EGFR mutation commonly occurred in the tumor tissue of the patients with TYMS mRNA low expression (Z=-2.604,P=0.009).Conclusion The TYMS mRNA expression in NSCLC tissue is correlated with the EGFR gene mutation,which can provide references for the drug selection aiming at the patients with different conditions.

OBJECTIVE:To establish a method for the content determination of saccharide in polysaccharides containing galact-uronic acid based on phenol-sulfuric acid method combined with correction factor method. METHODS:The determination condi-tion of phenol-sulfuric acid was optimized. A series of standard curves were drawn with glacturonic aid-glucose mixed control with different mass ratio. The content of saccharide in Codonopsis pilosula polysaccharides CPP1b samples containing galacturonic acid was determined. According to regression equation of galacturonic acid-glucose ratio of 0-100%,the correction factor was calculated by using C. pilosula polysaccharides CPP1b as the reference polysaccharide,and the results of content determination of saccharide were corrected. The rationality of this method was verified by using mixed monosaccharide control with same composition as C. pi-losula polysaccharides CPP1b. RESULTS:The correction factor of C. pilosula polysaccharide CPP1b to glucose was 3.33;in vali-dation test,the content of saccharide in mixed monosaccharide control with same composition as C. pilosula polysaccharides CPP1b was 103.47%. RSDs of precision and stability tests was <1%. The recoveries ranged 93.52%-107.35%(RSD=5.09%,n=6). CONCLUSIONS:The established method can accurately determine the content of saccharide in C. pilosula polysaccharides CPP1b containing galacturonic acid.

Objective To investigate the relationship between serum uric acid and traditional risk factors of coronary artery disease. Basic information was obtained by using questionnaire and measured for blood pressure , height ,weight. Venous blood was collected for detecting fasting plasma glucose ,serum lipid and uric acid. All data were analyzed with SPSS 20.0. Correlation analysis was used to assess the association of serum uric acid and traditional risk factors of coronary artery disease in patients with coronary artery disease. Results The mean value of serum uric acid in coronary artery disease is 6.3 mg/dL. The level of serum uric acid was significantly higher in male than that in female. Serum uric acid and body mass index(r = 0.137,P < 0.001),systolic blood pressure (r = 0.053,P = 0.025)and triglycerides(r = 0.188,P < 0.001)had a positive correlation. Serum uric acid was negatively correlated with high density lipoprotein cholesterol(r =-0.146,P < 0.001)and estimated glomerular filtration rate(r=-0.340,P<0.001). Serum uric acid was higher in patients with men,smoking,drinking,and chronic CAD. The serum uric acid level was lower in patients with exercise time more than 30 minutes than no movement or movement time less than 30 minutes. Conclusion Elevated serum uric acid were associated with body mass index,systolic blood pressure,triglyceride,high density lipoprotein cholesterol,estimated glomerular filtration rate,male,smoking,and chronic CAD.

OBJECTIVE To establish a new cell line that can stably express humanα2A-adrenoceptor (α2A- AR). METHODS Recombinant plasmid of α2A- AR with hygromycin B (Hygro) resistance (pcDNA3.1/Hygro-HA-α2A-AR)was stably transfected into Chinese hamster ovary(CHO)cells which had expressed protein kinase A catalytic subunits(PKAcat) with labeling of enhanced green fluorescent protein(EGFP)by a Lipofectamine based method. A single positive clone expressingα2A-AR was selected through cultivation in the presence of 200 mg · L-1 hygromycin B followed by PKA redistribution assay. The transcriptional expression ofα2A-AR was detected by quantitative real-time PCR(qRT-PCR). Time-resolved fluorescence resonance energy transfer immunoassay was used to identify the function of inhibiting cAMP accumulation of α2A-AR. RESULTS The CHO-PKAcat-α2A-AR cell line No.7 exhibited stable response in PKA redistribution assay. qRT-PCR analysis demonstrated that the high expression ofα2A-AR in the cell line remained stable after a few generations compared with CHO-PKAcat-EGFP cells (P<0.01). The cAMP accumulation caused by forskolin was significantly inhibited by α2A-AR agonist in CHO-PKAcat-α2A-AR cells(P<0.01). CONCLUSION CHO-PKAcat-α2A-AR cell line is constructed successfully, which provides an effective model for drug screening and studies of mechanisms.

Objective:Mutations in epidermal growth factor receptor (EGFR) can predict tumor response to tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). However, not all cases of NSCLC with EGFR mutations can respond well;thus, discovering the heterogeneity of NSCLC at the molecular level is necessary. This study aimed to determine the discrepancy in EGFR mutations in primary tumors, N2 lymph nodes, and plasma samples. Methods:Primary tumors, N2 lymph nodes, and plasma samples obtained from 49 patients with stageⅢA-N2 NSCLC were analyzed for EGFR mutations in exons 19 and 21 by using mutant-enriched liquidchip technology. Results:In 49 patients, we detected 18 (36.7%) EGFR mutations in primary tumors, 11 (22.4%) mutations in N2 lymph nodes, and 2 (4.1%) mutations in plasma samples. Eleven (22.4%) cases showed discordance in EGFR mutations between primary tu-mors and N2 lymph nodes. In nine cases, EGFR mutations were detected only in primary tumors, whereas EGFR mutations were de-tected only in N2 lymph nodes in two cases. In addition, EGFR mutations were detected in the plasma samples of two patients, who al-so carry mutations in their primary tumors. Conclusion:A considerable proportion of NSCLC cases showed discrepancy in EGFR muta-tions between primary tumors and N2 lymph nodes. In addition, the detection rate of EGFR mutations was lower in plasma samples obtained from patients with stage IIIA-N2 NSCLC. All of the results indicated tumor heterogeneity at the molecular level during metas-tasis, and this heterogeneity may have implications during treatment with TKIs.

Objective:This study aimed to achieve the early diagnosis and active treatment of adult hemophagocytic syndrome (HPS) and investigate the clinical characteristics and prognostic factors of this syndrome. Methods:A single-center retrospective analysis was performed to analyze clinical characteristics, laboratory findings, and survival data. Results:In 58 patients, the most common clinical manifestations were fever (100%) and splenomegaly (89.7%). The most common laboratory parameters were serum ferritin 500 g/L (100%) and peripheral cytopenia in two or more lineages (96.6%). platelet count, fibrinogen, and lactate dehydrogenase in the death group were significantly lower than in the survival group (P=0.000, 0.001, and 0.000). Survival analysis results showed that infections in the rheu-matological group exhibited good prognosis [the overall survival (OS) time was not reached in 190 d]. Patients with unexplained causes had moderate prognosis (OS time was 60 d);tumor-associated HPS patients had poor prognosis (the OS time was only 30 d). Univariate analysis results showed that patients with Fbg<1.5 g/L, PLT<40×109/L, and LDH≥2000 U/L also exhibited poor prognosis (P=0.000). Multivariate analysis results showed that PLT<40 × 109/L was an independent adverse factor (HR=6.472, 95%CI:1.526-26.065, P=0.011). Conclusion:HPS exhibits complex clinical manifestations and varied etiology. Patients with infection and rheumatism-related HPS had good prognosiss compared with those manifesting tumor-associated HPS. Fbg<1.5 g/L, PLT<40×109/L, and LDH≥2 000 U/L were the univariate factors that affected the survival time of patients. PLT<40×109/L is an independent adverse factor. These patients need systemic treatments as early as possible.

OBJECTIVE Toestablisharecombinantcelllinethatcanstablyexpressratpurinergic P2receptorP2X4(rP2X4R).METHODS ToconstructgreenfluorescentproteinandrP2X4recombinant plasmid (pEGFP-N1-rP2X4),lipofectamine was used to transfect pEGFP-N1-rP2X4 into human embry-onic kidney (HEK293)cells that were screened with G41 8 (1 g·L-1 ).The quantitative expression of rP2X4 receptor was verified by qRT-PCR and Western blotting analysis.Whole-cell patch clamp record-ing was used to investigate the function of the stably expressed rP2X4 receptor. RESULTS The sequence of plasmid pEGFP-N1-rP2X4 was verified by PubMed Blastn comparison.qRT-PCR and Western blotting analysis demonstrated that the expression of rP2X4 receptor in HEK293-pEGFP-N1-rP2X4 cell lines remained stable after 25 generations (P1 ,P3,P5,P10,P15,P20 and P25).Whole-cell patch clamp recording experiments showed that the rP2X4 receptor agonist,purine-5′-triphosphate (ATP,3.0 μmol·L-1 ),could activate rP2X4 receptors in HEK293-pEGFP-N1-rP2X4 cell lines.Specific activating current could be blocked by non-selective rP2X4 receptor antagonist TNP-ATP (30.0μmol·L-1).CONCLUSION rP2X4receptorisstablyexpressedinHEK293-pEGFP-N1-rP2X4cell line and maintains stable expression and function within 25 continuous generations.The establish ment of HEK293-pEGFP-N1-rP2X4 cell line can contribute to further investigations of the roles of rP2X4 receptors in neuropathic pain.

Schizophrenia, described as the worst disease affecting mankind, is a severe and disabling mental disorder. Schizophrenia is characterized by complicated symptoms and still lacks a diagnostic neuropathology, so developing schizophrenia animal models which have quantifiable measures tested in a similar fashion in both humans and animals will play a key role in new therapeutic approaches. According to the symptoms of cognitive impairment and emotional disorder, the N-methyl-d-aspartate (NMDA)-receptor antagonist MK-801 was applied to induce schizophrenia-like behavior in mice. Locomotor activity and prepulse inhibition (PPI) were selected as indices and the effect of clozapine was also investigated in this model. The results showed that compared with the normal group, MK-801-treated mice exhibited significantly increased locomotor activity and impaired PPI, and pre-exposure to clozapine could ameliorate the abnormality and make it back to normal level. These findings suggest that the model we established could be a useful tool for antipsychotic drug screening.