Objective To investigate the relation between serum asymmetric dimethylarginine (ADMA) level and endothelial function before and after living donor kidney transplantation in uremic patients.Methods A total of 38 renal transplant patients (21males and 17 females) and 36 healthy controls (20 males and 16 females) were enrolled. Plasma ADMA, symmetric dimethylarginine (SDMA), malondialdehyde (MDA), glutathione peroxidase (SeGSHPx), C-reactive protein (CRP) were mea-sured before transplantation and on Days 1, 3, 7, 14, and 28 posttransplantation. The brachial artery flow mediated dilatation (FMD) was studied before transplantation and on the 28th day after the transplantation. Results Serum levels of ADMA,SDMA,MDA and CRP were significantly increased, and the activities of nitric oxide (NO) and SeGSHPx were decreased in uremic patients compared with age matched healthy subjects (all P＜0.01). Serum levels of ADMA,SDMA,MDA and CRP decreased while the activities of NO and SeGSHPx increased significantly at the first day after the transplantation (all P＜0.01). The decrement of plasma SDMA normalized on the 28th day (P＞0.05). The FMD was lower in the patients than the control group (P＜0.01) and improved significantly on the 28th day of posttransplantation (P＜0.05). Serum levels of ADMA were positively correlated with MDA (r=0.412, P＜0.01;r=0.342,P＜0.01) and negatively correlated with the values of SeGSHPx (r=-0.345, P＜0.01;r=-0.315, P＜0.01) and FMD (r=-0.452,P＜0.01;r=-0.416,P＜0.01) both before and after kidney transplantation. Conclusion The level of serum ADMA is associated with endothelial function improvement in uremic patients both before and after kidney transplantation.

Objective To establish a new quality control standard for danshen capsules. Methods The qualitative identification of danshen capsules was characterized by ultraviolet fluorescence and thin-layer chromatography( TLC ). The contents of tanshinoneⅡA,cryptotanshinone,salvianolic acid B,danshensu and protocatechuic aldehyde in danshen capsules were determined by high performance liquid chromatography(HPLC)on a C18 column. The flow rate was 1 mL·min-1,and the column temperature was 35 ℃. Results The HPLC linear ranges of tanshinone ⅡA,cryptotanshinone,salvianolic acid B, danshensu and protocatechuic aldehyde were 2. 046-40. 92 μg · mL-1 ,1. 482 25 -59. 29 μg · mL-1 ,1. 502 55 -60. 102 μg·mL-1 ,11. 49-459. 582 μg·mL-1 ,and 0. 617 4-24. 696 μg·mL-1 ,respectively,and r values were 0. 999 9. The average recoveries were 99. 66%(RSD of 0. 91%)for tanshinoneⅡA,99. 26%(RSD of 0. 88%)for cryptotanshinone,99. 09%(RSD of 0. 76%)for salvianolic acid B,100. 51%(RSD of 0. 62%)for danshensu,and 100. 62%(RSD of 0. 82%)for protocatechuic aldehyde,respectively. The contents of the tanshinoneⅡA,cryptotanshinone,salvianolic acid B,danshensu showed a certain high level in the 3 batches of danshen capsule samples,but protocatechuic aldehyde was low by comparison. Conclusion The HPLC method is proven to be sensitive,accurate,repeatable,and can be used for quality control of the danshen capsules.

Background There is a growing interest in studying the relationship between intrinsic resistance and biofilms resistance to drugs. However, the relationship still remains unclear in the macroscopic bacterial growth. Our study is to illuminate the change of bacterial drug resistance of gyrA mutant and active efflux pump during the development of Pseudomonas aeruginosa (P. aeruginosa) biofilms. Methods The strains of type Ⅱ topoisomerase gene mutant (gyrA mutant) and multidrug resistance (MDR) efflux pump were clinical isolates and detected by polymerase chain reaction (PCR). The process of bacterial biofilms development was observed by scanning electron microscope. Triparental mating experiments were performed to transfer report gene of green fluorescent protein (GFP) into P. aeruginosa biofilms strains and followed by analysis of bacterial survival rate between intrinsic resistance and biofilms resistance.Results The fluorescent strains with pGFPuv could develop mature biofilms on Teflon surface. Before a period of 72 hours, the survival rate of biofilms bacteria and intrinsic resistance strains in ciprofloxacin solution was significantly different (P<0.05). The survival number of intrinsic resistance strains (gyrA mutation and active efflux pump) was illustriously higher than biofilm strain in the initial stage of biofilms development. After 72 hours incubation, there was no clearly difference between mutants and biofilms strains in the survival rate (P>0.05). The carbonyl cyanide m-chlorophenylhydrazone and azithromycin could significantly reduce the drug resistance of biofilm strains and efflux pump strains.Conclusions In the development of P. aeruginosa biofilms, the strains of gyrA mutation and MDR efflux could be conferred with new level of drug resistance. When co-cultured mutated strains with biofilm strains, biofilms may play a major role in bacterial resistance. But after 72 hours incubation (a mature biofilms had been developed), there was no clearly difference between the number of mutant strains and biofilm strains.

BACKGROUND: Propofol has been reported to protect vascular endothelial cells against oxidative stress. In this study we investigated its effect on hydrogen peroxide (H2O2)-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and examined the possible signaling pathways. METHODS: HUVECs were pretreated with propofol (1, 5, 25, and 50 microM) for 30 min and then co-incubated with 0.4 mM H2O2 for 4 h. Cell viability was assessed using a Cell Counting Kit-8. Cell apoptosis was analyzed using flow cytometry with annexin V/propidium iodide staining, and evaluated by quantifying caspase-3, Bax, and Bcl-2 expression levels. The expression levels of p38 mitogen activated protein kinase (MAPK), phosphorylated (p)-p38 MAPK, cJun-N-terminal kinases (JNK), phosphorylated (p)-JNK, Akt and phosphorylated Akt [(p)-Akt] (Ser473) were measured by western blotting. RESULTS: H2O2 treatment induced the activation of caspase-3, downregulated Bcl-2 expression, and up-regulated Bax expression, all of which were dose-dependently attenuated by propofol pretreatment. Furthermore, propofol significantly ameliorated H2O2-induced phosphorylation of p38 MAPK, JNK, and Akt in HUVECs. CONCLUSIONS: Propofol can protect HUVECs against H2O2-induced apoptosis via a mechanism that may involve p38 MAPK, JNK, and Akt signaling pathways.
Apoptosis* ; Blotting, Western ; Caspase 3 ; Cell Count ; Cell Survival ; Endothelial Cells ; Flow Cytometry ; Human Umbilical Vein Endothelial Cells* ; Humans* ; Hydrogen Peroxide ; Oxidative Stress ; p38 Mitogen-Activated Protein Kinases ; Phosphorylation ; Phosphotransferases ; Propofol* ; Protein Kinases

OBJECTIVETo investigate the expression of B7-H1 on peripheral blood mononuclear cells from patients with chronic HBV infection.

METHODSImmunofluorescent staining and flow cytometry assay were used to measure the expression of B7-H1 on peripheral blood CD3high T cells, CD19high B cells and CD14high monocytes from chronic HBV infected patients.

RESULTSNo significant difference was observed in B7-H1 expression on T cells and B cells between chronic HBV infected patients (CHB) and health controls (HC). B7-H1-expressing CD14high cells were significantly increased in chronic HBV-infected patients (19.17-/+11.64)% as compared with healthy controls [(7.30-/+5.49)%, P<0.01]. A significant positive correlation was found between B7-H1 expression on CD14high monocytes and serum ALT levels.

CONCLUSIONThere is no significant difference in B7-H1 expression on T cells and B cells between CHB patients and healthy subjects. B7-H1, which is up-regulated on monocytes from chronic HBV-infected patients, in positively correlated to serum ALT levels, and may play a role in the persistence of HBV infection.

Adult ; Aged ; Antigens, CD ; metabolism ; B-Lymphocytes ; immunology ; B7-H1 Antigen ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; immunology ; metabolism ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Monocytes ; immunology ; T-Lymphocytes ; immunology ; Young Adult

Objective To investigate the expression of Toll-like receptor(TLR) 9 of circulating plasmacytoid dendritic cell (pDC) and analyze the frequency and interferon (IFN)-?production of circulating pDC during hepatitis B virus(HBV) infection.Methods Peripheral blood was collected from 69 HBV-infected patients,including 14 cases of asymptomatie HBV infection,30 cases of chro- nic hepatitis B(CHB),25 cases of HBV-related liver cirrhosis,and 21 healthy blood donors as con- trols.Flow cytometry was used to analyze the frequency of circulating pDC and the mean fluorescence intensity(MFI) of TLR9.Fresh peripheral blood mononuclear cells(PBMC) were stimulated by CpG ODN 2216 for 24 h in vitro.IFN-?in the supernatant was measured using enzyme-linked immunosorbent assay(ELISA) to analyze the frequency and IFN-?production of pDC during HBV in- fection.Data were analyzed with SPSS 13.0 for windows.Results Compared with healthy controls (62.6?10.7),the MFI of TLR9 of patients with asymptomatic HBV infection,those of CHB and HBV-related cirrhosis were significantly reduced (P

OBJECTIVETo establish a method of fluorescence quantitative PCR to detect 21 trisomy syndrome.

METHODSAt first, using one pair of primer to simultaneously amplify different fragments of two highly homologous genes of the human liver-type phosphofructokinase located on chromosome 21 (PFKL-CH21) and the human muscle-type phosphofructokinase located on chromosome 1 (PFKM-CH1). Then, staining the PCR products of these homologous genes with SYBR Green I, comparing the fluorescence intensities of the bands after electrophoresis, and analyzing the data.

RESULTSThe relative fluorescence intensity ratios of PFKL-CH21/PFKM-CH1 in 21 trisomy syndrome and normal individuals were 1.58+/-0.17 (mean+/-SD) and 1.00+/-0.05 (mean+/-SD), respectively; the difference between the two groups was highly significant.

CONCLUSIONSYBR Green I fluorescence quantitative polymerase chain reaction is an acurate, rapid, safe and practical approach for the detection of 21 trisomy syndrome.

Child ; Child, Preschool ; Down Syndrome ; diagnosis ; genetics ; Female ; Fluorescence ; Humans ; Infant ; Infant, Newborn ; Male ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity

Objective Sperm screening is an essential step in IVF procedures. The swim-up method, an assay on sperm motility, is used clinically to select the ideal sperm for subsequent manipulation. However, additional parameters, including acrosome reaction capability, chemotaxis, and thermotaxis are also important indicators of mammalian sperm health. To monitor both sperm motility and chemotaxis simultaneously during sperm screening, we designed and constructed a microdevice comprising a straight channel connected with a bi-branch channel that mimics the mammalian female reproductive tract. Methods The width and length of the straight channel were optimized to select the motile sperm. Cumulus cells were selectively cultured in the bi-branch channel to generate a chemoattractant-forming chemical gradient. Sperm chemotaxis was represented by the ratio of the sperm swimming towards different branches. Results The percentage of motile sperm was improved from ( 58. 5 ± 3. 8 ) % to ( 82. 6±2.9)% by a straight channel 7 mm in length and 1 mm in width. About 10% of sperm were found chemotactically responsive in our experiment, which is consistent with previous studies. Conclusion The combined evaluation of both sperm motility and chemotaxis was achieved for the first time, and the motile and chemotactically responsive sperm can be easily enriched on a lab-on-a-chip device to improve IVF outcome.

BACKGROUND: Data on the mechanical ventilation (MV) characteristics and radiologic features for the cases with H7N9-induced ARDS were stil lacking. METHODS: We describe the MV characteristics and radiologic features of adult patients with ARDS due to microbiologically confirmed H7N9 admitted to our ICU over a 3-month period. RESULTS: Eight patients (mean age 57.38±16.75; 5 male) were diagnosed with H7N9 in the first quarter of 2014. All developed respiratory failure complicated by acute respiratory distress syndrome (ARDS), which required MV in ICU. The baseline APACHE II and SOFA score was 11.77±6.32 and 7.71±3.12. The overall CT scores of the patients was 247.68±34.28 and the range of CT scores was 196.3–294.7. The average MV days was 14.63±6.14, and 4 patients required additional rescue therapies for refractory hypoxemia. Despite these measures, 3 patients died. CONCLUSION: In H7N9-infected patients with ARDS, low tidal volume strategy was the conventional mode. RM as one of rescue therapies to refractory hypoxemia in these patients with serious architectural distortion and high CT scores, which could cause further lung damage, may induce bad outcomes and requires serious consideration. Prone ventilation may improve mortality, and should be performed at the early stage of the disease, not as a rescue therapy.

Objective:To investigate the clinical effectiveness of one-stage biplanar osteotomy correction of angulation and shortening deformity of the lower extremity after epiphyseal injury of the distal femur with a monorail external fixator.Methods:The data of 5 patients (2 males and 3 females) with angulation and shortening deformity in the lower extremity after epiphyseal injury of the distal femur were retrospectively analyzed.The patients underwent monorail external fixator assisted one-stage osteotomy correction of the distal femur and distraction osteogenesis of the middle and upper femur in Zhengzhou Orthopeadics Hospital from May 2017 to December 2019.The mean age was 13.6 years old (range: 10 to 17 years old). The affected limbs were shortened by 5.1 cm on average (range: 3.9 to 6.5 cm). The average angulation deformity of the distal femur was 24.9° (range: 17.0°to 30.5°). The mechanical lateral distal femoral angle (mLDFA), the mechanical posterior distal femoral angle (mPDFA), the mechanical axis deviation (MAD), the range of motion (ROM) of the knee, and the length of the lower limbs before surgery and at the final follow-up were measured and analyzed.Results:All patients were followed up for 22 months on average (range: 15 to 32 months). For all the 5 patients, the mechanical axis was well realigned, mLDFA, mPDFA, and MAD returned to normal range, and the length of the affected limb achieved the goal as planned before the surgery.Besides, the affected limbs were lengthened by 5.6 cm on average (range: 3.9 to 8.0 cm), and the median healing index was 35.6 d/cm (range: 29.0 to 45.0 d/cm). The bone callus in the distraction area and the osteotomy end were well healed at the final follow-up, as indicated by the X-ray results.At the end of the distraction period, the flexion ROM of the knee in all patients reached basically 90°.By the final follow-up, all patients had a normal knee ROM.No vascular or nerve injury, dislocation of hips or knee joints, re-fracture after disassembly, deep infection and other complications were observed in all patients.Conclusions:One-stage biplanar osteotomy correction of angulation and shortening deformity of the lower extremity after epiphyseal injury of the distal femur with a monorail external fixator is safe and feasible.The method requires no multiple operations and improves the tolerance of patients during the treatment period.