Objective To evaluate the application value of glycerol blood chocolate soup in the bacteria preservation of Haemophilus influenzae.Methods Haemophilus influenzae strains ATCC10211,ATCC49247 and ATCC49766 were saved into glycerol blood chocolate soup,then it was stored in refrigerator at below -35℃.The preserved strains from the refrigerator at 1 month,3 months,6 months,1 year,2 years were taken out,and dissolved it at 35℃ for 5min,and then immediately transferred it to chocolate agar plate,placed it in the 35℃,CO2 environment overnight.Observed the survival of strains,colony morphology and whether the demand factor test had change.Saved the standard strains ATCC49247 and ATCC49766 2 years for the paper dispersion method drug sensitive test.Results After 2 years of observation,3 strains of Haemophilus influenzae strains grew well in the survival rate of 100%,the colony was colorless,transparent and flat moist,it could be observed gram small negative bacilli by Gram staining, blood agar satellite test was positive,no hemolysis,Columbia satellite test was negative.All traits were the characteristics of Haemophilus influenzae.The drug sensitivity characteristics of ATCC49247,ATCC49766 standard strains were still in line with the requirements of CLSI.Conclusion Glycerol blood chocolate soup is suit for Haemophilus influenzae bacteria preservation.

Objective To evaluate the quality and application value of improved sheep blood chocolate medium.Methods The ATCC 10211 of Haemophilus influenzae was inoculated into the modified medium and unmodified medium,the average growth index(GI)of Haemophilus influenzae in two kinds of culture medium was compared.Based on 352 selected qualified sputum specimens for detection of Haemophilus influenzae,the positive isolation rate of medium was compared between the two groups.Results GI value of the traditional blood chocolate culture medium was (3.69 ±0.58),which was significantly lower than (15.08 ±1.34)of the improved sheep blood chocolate culture medium,the difference was significant (t =25.31,P <0.01 ).352 sputum specimens in the improved sheep blood chocolate culture medium,Haemophilus influenzae detected in 41 cases,the positive rate was 11.65%.And 352 sputum specimens in traditional sheep blood chocolate culture,Haemophilus influenzae detected in 18 cases,the positive rate was 5.54%.There was significant differenceof Haemophilus influenzae in separation based on 2 kinds of chocolate culture medium (χ2 =21.04,P <0.05).Conclusion Haemophilus influenzae significantly improved sheep blood colony in chocolate culture medium,grows well,easily to be identified,which helps to detect sputum specimens of Haemophilus influenzae.

Diabetic nephropathy (DN) is one of the most common microvascular complications in diabetes mellitus patients and is characterized by thickened glomerular basement membrane,increased extracellular matrix formation,and podocyte loss.These phenomena lead to proteinuria and altered glomerular filtration rate,that is,the rate initially increases but progressively decreases.DN has become the leading cause of end-stage renal disease.Its prevalence shows a rapid growth trend and causes heavy social and economic burden in many countries.However,this disease is multifactorial,and its mechanism is poorly understood due to the complex pathogenesis of DN.In this review,we highlight the new molecular insights about the pathogenesis of DN from the aspects of immune inflammation response,epithelial-mesenchymal transition,apoptosis and mitochondrial damage,epigenetics,and podocyte-endothelial communication.This work offers groundwork for understanding the initiation and progression of DN,as well as provides ideas for developing new prevention and treatment measures.

Objective To evaluate the quality of domestic hepatitis C diagnostic reagents objectively,and to build up the quality control systems for assessment of hepatitis Cdiagnostic reagents.Methods4080 serum samples from blood donors were collected and detected with EIA kits.146anti-HCV positive and negative samples were selected and tested repeatedly by two different imported ( Murex and Ortho) and domestic anti-HCV EIA kits(InTec,ZHONGSHAN BIO-TECH,WANTAI and KHB),then confirmed by CHIRON RIBA HCV 3.0 and PCR qualitative reagents.The samples were tested by nucleic acid quantitative assay and the RNA positive samples were detected by genotyping reagents.ResultsThe quality control systems of diagnostic reagents of anti-HCV and HCV RNA were constructed.Each quality control system was consisted of 50 samples,including 20 anti-HCV/HCV RNA positive,20 anti-HCV/HCV RNA negative and 10 diluted specimens for sensitivity evaluation.The positive samples with dominant HCV genotypes in China contained strong,moderate and weak positive samples.The negative samples involved those S/CO value ( signal-to-cutoff ratios ) close to threshold.Conclusion The quality control systems established in this study are suitable for assessment of the new and improved domestic hepatitis C diagnostic reagents.

This study is designed to investigate the effects of chinfloxacin hydrochloride (CFX) on the kinetics of HERG K+ channel. Whole cell patch clamp technique was used to record HERG K+ currents from HEK293 cells transiently transfected with cgi-HERG-GFP plasmids and channel kinetics were assessed in the absence and presence of CFX and moxifloxacin hydrochloride (MOX). Results demonstrated that the open state of HERG K+ channel was inhibited by CFX in a concentration- and time-dependent manner, with an IC50 of 162.1 +/- 14.2 micromol x L(-1), two folds higher than its positive control MOX. But there were no significant effects on channel kinetics. In addition, the inhibitory effect of CFX on HERG was enhanced when cells were subjected to altered extracellular K+ concentration.

Objective To explore the potential mechanisms of mesenchymal stem cell (MSC) therapy in ischemia/reperfusion injury (IRI)-induced acute kidney injury (AKI). Methods Forty-five C57/BL6 male mice were randomly divided into three groups: sham group, IRI group, and IRI+MSCs group, with 15 mice in each group. The IRI-induced AKI model in mice was reproduced by clamping both renal pedicles for 35 minutes. In the sham group, both kidneys were exposed, but their pedicles were not clamped. Six hours after reperfusion, mice in IRI+MSCs group received 100 μL of MSCs (1×104 /μL) isolated from the bone marrow from C57/BL6 mice via tail vein, while the mice in the IRI group received same amount of normal saline. Blood samples were harvested at 48 hours after reperfusion, and levels of serum creatinine (SCr) and blood urea nitrogen (BUN) were determined. The changes in renal pathology were observed by microscopy with PAS staining, and the tubular injury and acute tubular necrosis (ATN) scores were calculated. The number of leukocytes (CD45+) infiltrated in kidney at 24 hours and 72 hours after reperfusion was measured with flow cytometry. The number of neutrophils (Ly-6G+) and macrophages (F4/80+) infiltrated in kidneys at 24 hours and 72 hours after reperfusion was determined by immunofluorescence. Results There was significant increase in the related parameters in IRI group compared with those of sham group. The levels of SCr (μmol/L) and BUN (mmol/L) were 180.3±8.8 vs. 9.7±3.5, and 1 121.1±8.3 vs. 9.4±2.3, both P < 0.01. The score of tubular injury was 4.80±0.55 vs. 0 at 48 hours after reperfusion. The quantity of leukocyte (CD45+) infiltration in kidney at 24 hours and 72 hours after reperfusion was increased (×105 cells/g: 60.50±2.56 vs. 19.46±4.83, 42.00±1.87 vs. 14.70±3.74, both P < 0.01), and the number of neutrophils (Ly-6G+) and macrophages (F4/80+) infiltrated in kidney at 24 hours and 72 hours after reperfusion was also increase although the number of leukocytes infiltrated in kidney was significantly lower at 72 hours after reperfusion than that at 24 hours. There was significant lowering of the levels of SCr and BUN [SCr (μmol/L): 99.0±8.0 vs. 180.3±8.8, BUN (mmol/L): 84.5±7.6 vs. 112.1±8.3, both P < 0.01] in IRI+MSCs group, compared to IRI group. For the degree of tubular necrosis in two groups, the tubular injury scores were 2.60±0.55 vs. 4.80±0.55 (P < 0.05). The number of leukocytes infiltrated in kidney at 24 hours and 72 hours after reperfusion (×105 cells/g) were 24.20±4.53 vs. 60.50±2.56, 31.70±3.15 vs. 42.00±1.87 (both P < 0.01). The number of neutrophils was lowered despite (the number of macrophages was increased). However, the number of infiltrated leukocytes was significantly more in IRI+MSCs group at 72 hours than that at 24 hours (×105 cells/g: 31.70±3.15 vs. 24.20±4.53, P < 0.05). Conclusion MSCs could protect against IRI induced AKI by reducing the total number of leuckocytes, especially that of the neutrophils infiltrating into ischemic kidney and by recruiting macrophages into ischemic kidney.

Objective To explore the efficacy and safety of fondaparinux combined with tirofiban in patients with high risk unstable angina (UA) undergoing complex percutaneous coronary intervention (PCI).Methods A total of 389 patients were enrolled and randomized into two groups receiving either fondaparinux with tirofiban or enoxaparin with tirofiban.Bleeding,thrombosis and main adverse cardiovascular events (MACE) were compared between the two groups during hospitalization,at week 2 and week 4 after discharge.Results No severe bleeding was observed during hospitalization in the both groups,while lower rate of mild and minor bleeding was shown in the fondaparinux group (0 vs 1.5％ and 18.2％ vs 34.5％,P =0.04 and P ＜0.001 respectively).No difference was found between the two groups in the rate of MACE during hospitalization,at week 2 and week 4 weeks after discharge.The rates of death,recurrent myocardial infarction,refractory myocardial ischemia and target vessel revascularization were 0.5％ vs 1.0％,0.5％ vs 1.0％,1.6％ vs 1.0％ and 2.1％ vs 1.5％ during hospitalization;0 vs0,1.0％ vs 0.5％,1.0％ vs 1.5％,0.5％ vs 1.0％ at week2 after discharge; 0.5％ vs0.5％,0.5％ vs0.5％,2.6％ vs 2.0％,0 vs 0.5％ at week 4 after discharge (all P values ＞ 0.05).Conclusion The combination therapy of fondaparinux and tirofiban is of good safety and efficacy in high risk UA patients undergoing complex PCI.

Objective To explore the role of clinical application of enteral nutrition sequential therapy in early enteral nutrition support by comparison with enteral nutrition non-sequential therapy in critically ill patients with cerebrovascular diseases.Methods A total of 62 patients were randomly (random number) divided into sequential group and conventional (non-sequential) grouThe comparisons of tolerance for enteral nutrition support,levels of prealbumin,the mechanically ventilated time and mortality rate in 28-day between two groups were carried out.Results The tolerance of sequential group was superior to that of conventional group (P＜0.05).The higher level of prealbumin and the shorter mechanical ventilation time were observed in sequential group compared with conventional group (P＜0.01).Compared with conventional group,the patients in sequential group had lower mortality rate in 28 days (P＜0.05).Conclusions Sequential therapy is beneficial to the implementation of early enteral nutrition support in patients with severe cerebrovascular disease,reducing the nutritional adverse events,and improves the prognosis.

Recruitment maneuver(RM) refers to the process of reopening collapsed alveoli through transient lung inflations with high pressure during the mechanical ventilation in order to improve arterial oxygenation and respiratory mechanics,attenuate ventilator-induced lung injury.At present,a large number of animal experiments and clinical studies have focused on the effect of RM or factors affecting RM in acute respiratory distress syndrome.Pulmonary edema is an important pathophysiological feature of acute respiratory distress syndrome,so the effect of RM on pulmonary edema is worthy of our attention.Here is to make a review of the current progress.

Objective:To explore the effect of augmented renal clearance(ARC)on 24-hour area under the concentration-time curve to minimum inhibitory concentration ratio(AUC 24/MIC)of vancomycin and prognosis in critical children, thus to provide proposal for individual dosage regimen. Methods:Sixty-five critical children treated with vancomycin, who suffered from sepsis/septic shock, were brought into this retrospective cohort study.According to estimate glomerular filtration rate, these children were divided into ARC group ( n=27) and normal group ( n=38). The influencing factor of AUC 24/MIC of vancomycin and therapy prognosis for two groups were detected and analyzed. Results:There were no significant differences between two groups in basic setting (age, sex, weight), scores of pediatric sequential organ failure assessment and pediatric risk of mortality Ⅲ, infection markers (C-reactive protein and procalcitonin), glutamic-pyruvic transaminase, hypoproteinemia, usage of diuretic and vasoactive agent( P>0.05). The patients from ARC group showed lower levels than those from normal group in AUC 24/MIC of vancomycin[375.2(300.8, 489.4) vs. 443.6(412.3, 593.2), Z=2.263, P=0.024] and it′s target achievement ratio (TAR)(40.7% vs. 76.3%, χ2=8.440, P=0.005). When usage of diuretic and vasoactive agent, the AUC 24/MIC of ARC group was lower than that of normal group( P<0.05). But there was no significant difference between ARC group and normal group regarding hypoproteinemia( P>0.05). The days of body temperature steady at least 48 hours[7.0(5.5, 9.0)d vs. 6.0(5.0, 8.0)d], the length of hospital stay[39.0(21.0, 58.0)d vs. 20.5(16.0, 28.0)d], the length of PICU stay[14.0(9.0, 31.5)d vs. 10.0(5.0, 15.0)d] were longer than those in normal group( P<0.05). There were no significant differences between ARC group and normal group regarding days of ventilation and infectious markers decreased at least 50%, as well as 28-days mortality( P>0.05). The multivariable analysis showed that the presence of ARC, hypoproteinemia, use of diuretics and vasoactive agent were significantly associated with AUC 24/MIC of vancomycin( P<0.05). Conclusion:ARC may down regulate levels of AUC 24/MIC and TAR of vancomycin.During ARC period, the usage of diuretic and vasoactive agent could affect the AUC 24/MIC of vancomycin.Individual dosage regimen should be employed for critical children suffered with ARC.