Continuous blood purification therapy, one of the most popular multiple organ protection and life support technology, has been widely used in the treatment of critical illness.After the initial introduction of the concept of dialytrauma or CRRT trauma in 2012, the related complications are getting more and more attention.The loss of heat caused by CBP may lead to hypothermia.Howev-er, to critically ill patients, controversy still reigns on the influence of this issue.There is no guideline developed through clinical prac-tice to define such issue.This article reviews the methods as well as results of related clinical studies to discuss how to manage the hy-pothermia during CBP.

[Abstract ] Objective To understand the identification and in vitro antifungal susceptibility of Penicillium marneffei(PM)in yeast phase,and guide clinic antifungal application.Methods Strains isolated from blood and bone marrow of 23 patients infected with PM in a hospital between 2009 and 2016 were collected,colony morpholo-gy of PM in yeast phase was observed,susceptibility to itraconazole,voriconazole,amphotericin B,and fluconazole were detected with E-test method.Results Colony morphology of PM were as follows:direct microscopic examina-tion of Wright’s staining of tissue specimens found visible oval or round spore with apparent septum,and mainly lo-cated in macrophage;Gram staining of blood culture specimens found that strains were with bulbous and slightly curved ends,occasionally branched and with septum. PM was dimorphic fungi,presented mycelium at 28° C,pro-duced red pigment and diffused into medium;PM presented yeast form at 35° C,there were typical colony morpholo-gy. Minimum inhibitory concentrations (MICs)of itraconazole,voriconazole,amphotericin B,and fluconazole to PM in yeast phase were 0.002-0.016,0.012-0.125,0.002-0.500,and 0.500-16.000μg/mL respectively. Conclusion Typical colony morphology and fungal spore of PM in bone marrow and peripheral blood are important features for identification. PM is most susceptible to itraconazole,followed by voriconazole and amphotericin B, while fluconazole is less susceptible.

AIM:To investigate morphologic and functional changes of small intestinal mucosa and proliferating cell nuclear antigen in postoperative portal hypertension patients with single or combined administration of Gln and rhGH.METHODS:Twenty-nine portal hypertension patients with surgical treatment were prospectively randomized to four groups as follows:① Gln group(n=6);② rhGH group(n=8);③ Gln+rhGH group(n=7)and ④ control group(n=8).A standard solution for TPN was given three days after operation for a week.The concentration ratio of urinary lactulose and mannitol(L/M),the villus height and crypt depth and PCNA index of small intestinal mucosa were compared.RESULTS:A week after TPN postoperation,the increased ratios of L/M in Gln+rhGH group were less than those in control group(P0.05).CONCLUSION:This study suggest that Gln together with rhGH reduce the intestinal permeability and protect the mucosa integrality in postoperative portal hypertension patients,but not in single treatment.

Objective To explore the possible mechanism of miR 34a/Bcl-2 apoptotic pathway in the mouse model of age-related hearing loss.Methods A C57BL/6 mouse model of age-related hearing loss was conducted,and 4-week-old and 12-month-old mice were considered as the objects.The auditory brainstem response (ABR) was used to test the hearing function.The TdT-mediated dUTP nick end labeling (TUNEL) was used to observe the apoptosis of neuron in auditory cortex.The mRNA and protein levels of miR-34a,Bcl-2 and caspase 3 were detected by real-time PCR and Western bloting,respectively.Results The ABR showed that the hearing threshold level at 4,8,16,32 kHz were higher in 12-month-old mice than in 4-week-old mice [(80.0±2.5) dBHL vs.(32.0 ±4.5) dBHL,(74.0±3.5) dBHL vs.(51.0±1.2) dBHL,(86.0±4.6) dBHL vs.(51.0±3.5) dBHL,(87.0±6.6) dBHL vs.(56.0±1.5) dBHL,all P＜0.05].Compared with 4 week-old mice,the total number of neurons in auditory cortex was decreased,the number of apoptosis neurons was increased,the expressions of miR-34a (t=6.02,P=0.001),Bax (t=6.51,P=0.012) and Caspase 3 (P=0.023) rised,and the expression of Bcl-2 (t=7.12,P=0.032) declined in 12 month-old mice.Conclusions The activation of miR-34a/Bcl-2 apoptotic pathway may be one of the mechanisms of age-related hearing loss.

Objective To investigate the necessity of dilution in samples with high concentrations of D-Dimer and optimum dilution multiple.Methods Quality control products and calibration were detected by using Sysmex CS5100 for precision evaluation,including within-batch and between-run precision.Calibration were detected for validation of linear range and clinical reportable.Samples with D-Dimer<5 mg/L and fibrinogen degradation products(FDP)<20 μg/mL were serially diluted and detected to calculate recovery rate.Samples with D-Dimer>5 mg/L and FDP>20 μg/mL were also serially diluted and detected to calculated recovery rate.Results Within-batch and between-run coefficients of variation were both less than 3%.Within the scope of 0.207-5.170 mg/L,the linear distribution was fine.The clinical reportable range was 0.207-165.440 mg/L.For samples with D-Dimer<5 mg/L and FDP<20 μg/mL,no antigen excess phenomenon was found,and gradient dilution was not necessary.For samples with D-Dimer>5 mg/L and FDP>20 μg/mL,there was obvious antigen excess phenomenon,and gradient dilution was required.Conclusion For samples with D-Dimer>5 mg/L and FDP>20 μg/mL,dilution should be performed to ensure the accuracy of detected results.

Objective: To compare two methods (microbial assay and radioimmunoassay) for measuring plasma folate concentrations, and to examine the relationship between plasma folate levels, and alcohol consumption, tobacco use and body mass index, and the risk of hyperhomocysteinemia in China. Methods: We used a microtiter plate microbial assay and a radioimmunoassay to measure the folate concentration in 88 plasma samples. After comparing the results of these two methods and fitting a regression line, we examined the geographical, seasonal, and gender differences in folate concentration of plasma collected from 2 422 adults in south and north areas in China, and evaluated the association of plasma folate concentration, with alcohol consumption, cigarette smoking, and body mass index, and with the risk of hyperhomocysteinemia, using the data from the two assays. Results: The data from the two assays had a linear relationship ( r =0.879, P =0.000); the regression was Y =0.683 X +0.308 (where X and Y were nature logarithmic transformations of plasma folate by microbial assay and radioimmunoassay, respectively); however, the mean plasma folate levels by microbial assay were much higher than those obtained by radioimmunoassay. Both data sets showed similar plasma folate distributions among Chinese adults, associations with other risk factors, and the risk of hyperhomocysteinemia. We estimated that 19.9% of the Southerners and 67.1% of the Northerners had plasma folate concentrations by radioimmunoassay lower than the 6.8 nmol/L used to define plasma folate deficiency. Conclusion: There is a linear relationship between plasma folate levels determined by microbial assay and radioimmunoassay, but because of the different levels obtained in the two assays, it is difficult to use the microbial assay results to evaluate folate status at this time. The use of 10.5 nmol/L as a cut off for plasma folate deficiency by microbial assay needs further study.

Objective To evaluate the Carryover between the chemistry and immunoassay in Beckman Coulter Laboratory Au-tomation System and decide to whether sharing samples for testing between chemistry and immunoassay systems or not. Methods According to a certain order,high concentration samples and low concentration samples of HCG with different sample volume (500 μl,2 000 μl)were tested on Beckman AU5421 automatic biochemical analyzer.The HCG of low concen-tration samples were then tested to evaluate the carryover between the chemistry and immunoassay and explored the correc-tive procedure to deal with the carryover by increasing special cleaning process of beckman AU5421 automatic biochemistry analyzer.Results Under different sample volume,the carryover in a single module and as a whole of the beckman AU5421 automatic biochemistry analyzer were 5.44,15.47,23.51 and 45.96 ppm respectively (t=14.553,P <0.001;t=5.527,P =0.005;t=3.985,P =0.016;t=20.457,P <0.001).By increasing special cleaning process the carryover of 0.22 ppm was detected in 500 μl sample volume of the beckman AU5421 automatic biochemistry analyzer as a whole.Conclusion The car-ryover between the chemistry and immunoassay in Beckman Coulter Laboratory Automation System could been sovled by in-creasing special cleaning process of beckman AU5421 automatic biochemistry analyzer.

Background Choroidal melanoma(CM)is a common form of primary ocular cancer in adults.It is reported that indomethacin has inhibitory effect on many tumor in vitro and in vivo,but whether it can inhibit the growth of CM has not been published. Objective This study was to investigate the anti-tumor activity of indomethacin on the growth of human CM OCM-1 cell xenografts in nude mice. Methods OCM-1 cells were subcutaneously implanted on 24 SPF female BALB/C.nu/nu nude mice to establish ectopic models of human CM.The nude mice with the tumor 5 mm were randomly divided into 4 groups:untreated group,normal saline solution(NS) group,indomethacin 1 ms/kg group,indomethacin 2 ms/kg group.The 1 mS/kg or 2 ms/kg indomethacin was intraperitoneally injected for 14 consecutive days in indomethacin 1 ms/kg group and indomethacin 2 me/kg group respectively.and 0.2 ml of 2%NS-DMSO was used at a same way in the NS group.No any agent was used as the untreated group.The volume and weight of implanted tumor as well as inhibitory rates of indomethaein on tumor were calculated.The expression of ki67 and survivin proteins were measured with immunohistochemistry,and the expression of survivin mRNA in CM was assessed by RT-PCR. ResuIts The tumor of indomethacin treatment group was reduced in volume and weight with a significant difference between treatment group and control group as well as indomethacin 1 ms/ks group and indomethacin 2 ms/kg group(P＜0.05).The inhibitory rate of indomethacin 1 ms/kg and 2 ms/kg for tumor was 22.86%,48.00%respectively.The prolifiration index (PI)of ki67 in these 4 groups were (76.73±3.34)%,(73.30±2.95)%,(55.97±2.24)%,(32.87±2.91)%respectively,and significant difference was found in PI between indomethacin 2 mg/kg group and untreated group or NS group(P＜0.05),but there was not significant difference between indomethacin 1 mg/kg and 2 ms/kg group(P＞0.05).Compared with the control group,the indomethacin treatment groups showed the decreased expression of survivin protein and mRNA,and significant difference was found between indomethaein 2 ms/kg group and untreated group or NS group(P＜0.05),however,no significant difference was found between indomethacin 1 mg/kg and 2 mg/kg group(P＞0.05). Conclusion Indomethacin inhibits the growth of CM in nude mice through inhibiting the expression of survivin in the tumor and accelerating cell apoptosis and inhibiting tumor cell proliferation.

OBJECTIVETo investigate the correlation between matrix metalloproteinase 9 (MMP-9) expression and tumor invasion and metastasis as well, and to explore the potential application of controlled expression of target gene in tumor gene therapy.

METHODSOne self-contained tetracycline-regulated retroviral vector containing anti-sense cDNA of MMP-9 was constructed and transfected into a metastatic human melanoma cell line WM451 which expressed a high level of MMP-9. Techniques such as growth rate measurment, MTT assay, (3)H-thymidine incorporation, colony forming ability in soft agar, invasion assay in Boyden chamber, as well as zymography and Western blot were applied to analyze the expression of MMPs and behaviors of tumor cells in vitro before and after gene transfection. Tumorigenecity and spontaneous metastasis were tested in nude mice.

RESULTSIn the presence of exogenous tetracycline, the transfected antisense MMP-9 did not affect the endogenous level of MMP-9 in WM451 cells, and showed no significant changes in cell behaviors in comparison with that of the vector-transfected control cells. Nevertheless, withdrawal of tetracycline from the medium caused a significant down-regulation of expression and activity of MMP-9. The capacity of cell growth in vitro, colony forming ability in soft agar, invasion through Matrigel all were inhibited remarkably when compared with the controls. Spontaneous metastasis in nude mice was significantly inhibited.

CONCLUSIONSTransfection of anti-sense MMP-9 can down-regulate the invasion and metastasis of melanoma cells both in vitro and in vivo, further clarifying the important role of MMP-9 in tumor progression.

Animals ; Cell Division ; Cell Line, Tumor ; DNA, Antisense ; DNA, Complementary ; genetics ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Genetic Vectors ; Humans ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Melanoma ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; Retroviridae ; genetics ; Tetracycline ; pharmacology ; Transfection

Antibiotic-associated diarrhea(AAD)in children is a type of diarrhea that occurs after the use of antibiotics in children,and its pathogenesis is closely related to the intestinal flora.The medication of antibiotics can affect the metabolic function of the intestinal flora and the immune function of the body,and then leads to the occurrence of AAD.In the view of Chinese medicine,AAD in children is mainly involved the spleen,and the etiology of the disease is due to the weakness of the spleen and stomach of the body constitution together with the attack of the pestilential pathogen and the accumulation of drug toxin.The pathogenesis of ADD in children is characterized by spleen deficiency with predominant dampness,deficiency of spleen qi,and insufficiency of spleen yang.Spleen deficiency is the root cause of pediatric AAD,and spleen and intestinal flora have commonality,so the treatment of pediatric AAD can be performed from the perspective of the spleen.The treatment of pediatric ADD from the spleen follows the principle of strengthening and activating the spleen,and the regulation of the spleen for achieving the purpose of treating the disease from the root can be achieved by the methods of strengthening spleen and draining dampness,strengthening spleen and replenishing qi,and strengthening spleen and warming yang separately with the fundamental prescriptions of Shenlin Baizhu Powder,Sijunzi Decoction,and Fuzi Lizhong Pills.