(Pro)renin receptor(PRR)was organically identified as a specific receptor for prorenin and renin to regulate the activity of renin-angiotensin system(RAS). Increasing evidence suggests that PRR plays a multitude functions in a RAS-dependent and independent manner. The extracellular domain of PRR is cleaved by furin or ADAM19 to produce a 28-ku soluble PRR(sPRR) while the intracellular domain,M8.9,is a subunit of vacuolar H+-ATPase that mediates H+transport. PRR is critically involved in embryogenesis in low vertebrates and mammals. In recent years ,a significant progress has been made in identifying the physiological and pathophysiological functions of renal PRR. PRR has been identified as an important regulator of urine concentrating capability mostly due to its ability to upregulate to aquaporin-2(AQP2)in the collecting duct and Na+,K+,2Cl-cotransporter(NKCC2)in the thick ascending limb. PRR also promotes K+ secretion in response to K+loading through extra-adrenal aldosterone(Aldo)produc?tion. Overactivation of renal PRR contributes to the pathogenesis of hypertension induced by angiotensin Ⅱ infusion and fructose/salt. Overall,PRR has emerged as a new regulator of physiological and pathophysiological processes in the kidney.

A molecularly imprinted film ( MIF) was prepared on the surface plasmon resonance ( SPR) sensor chip for the detection of testosterone. The nanometer imprinted film was synthesized by surface grafting approach. First, the gold sensor chip was modified with an iniferter of benzyl diethyldithiocarbamate to create the “grafting from” polymeric surface. Grafting of the MIF onto the SPR chip was subsequently achieved through ultraviolet light-initiated copolymerization of methacrylic acid ( functional monomer ) and ethylene glycol dimethacrylate ( crosslinker ) in the presence of testosterone as a model template. The template molecules were then removed to form a MIF with specific recognition sites for testosterone. With this iniferter technique , self-aggregation in the reaction solution phase was avoided and grafting polymerization was confined to the exterior of the chip surface. The grafting process was implemented by in situ monitoring with SPR, which permitted the thickness of the film to be easily and strictly controlled. The chip surface modified with a testosterone-imprinted film was characterized by the methods of polarization modulation infrared reflection absorption spectroscopy ( PM-IRRAS ) and atomic force microscopy ( AFM ) . According to the results, testosterone-imprinted film modification of the SPR chip was achieved by the distribution of numerous homogeneous, nanoscale holes on the surface. The testosterone-imprinted film was applied on a SPR sensor chip for the detection of testosterone in the range of 2 . 5 í 10-16 to 2 . 5 í 10-6 mol/L in acetonitrile with the lowest determining concentration of 2. 5 í 10-16 mol/L. Adsorption experiments showed that there were two kinds of binding sites in the MIF, and the linear correlations between the changes in reflectivity and the concentration of testosterone were y=19. 69 + 1. 21x(R2=0. 9913) and y=11. 5 + 0. 45x(R2=0. 9895), respectively. Control experiments utilizing estradiol, estriol, and progesterone as analogues showed impressive selectivity and specificity for testosterone determination. The testosterone-imprinted film reproducibility was evaluated with five cycles of rinsing-rebinding and the RSD was 16 . 8% and 11 . 2% for the SPR angle changes in rinsing and rebinding respectively, demonstrating that the MIF-modified SPR sensor had good reproducibility and repeatability. Finally, the SPR sensor was successfully used to determine testosterone in artificial urine samples with recoveries from 85 . 2% to 92 . 8%.

Objective To investigate the clinical characteristics of elderly patients with multiple myeloma and their prognostic factors.Methods 100 multiple myeloma patients with age≥ 60 years and 100 multiple myeloma patients with age <60 years who admitted in our hospital from December 2007 to December 2015 were collected as research subjects.100 patients with age ≥60 years were divided into elderly multiple myeloma group,100 cases with aged <60 years were divided into non -elderly multiple myeloma group.The clinical data and laboratory results of two groups were compared,the prognosis factors in elderly patients with multiple myeloma were analyzed.Results The incidence rate of ISS stage Ⅰ -Ⅱ in elderly multiple myeloma group (45.0%)was lower than non -elderly multiple myeloma group (60.0%),the incidence rate of stage Ⅲ was higher than non -elderly multiple myeloma group (χ2 =4.511,P <0.05).The infection incidence of elderly multiple myeloma group(30.0%)was higher than non -elderly multiple myeloma group(15.0%)(χ2 =10.452,P <0.05 ).The hemoglobin,serum albumin contents of elderly multiple myeloma group [(83.7 ±19.8)g/L,(27.89 ±6.87)g/L]were less than non -elderly multiple myeloma group[(92.1 ±22.5)g/L,(33.15 ±7.69)g/L](t =4.297,4.426,all P <0.05).The calcium content,the propor-tion of bone marrow plasma cells,serum creatinine and blood β-microspheres protein levels of elderly multiple mye-loma group [(2.51 ±0.41)mmol/L,(39.43 ±18.64)%,(182.24 ±125.47)μmol/L,(9.02 ±6.24)mg/L]were higher than non -elderly multiple myeloma group [(2.36 ±0.48)mmol/L,(37.45 ±19.86)%,(143.25 ± 116.43)μmol/L,(5.87 ±3.41)mg/L](t =5.945,4.196,4.375,4.264,all P <0.05).The median survival time of elderly multiple myeloma patients were significantly correlated with the patients'age,the proportion of plasma cells in bone marrow,blood β2 -microglobulin,albumin,and ISS staging (χ2 =4.125,3.254,8.542,5.748,9.244,all P <0.05).Conclusion The condition of elderly myeloma patients is more serious,age,proportion of plasma cells in bone marrow,blood β2 -microglobulin,albumin and ISS stage affect myeloma patients prognosis.

Objective To investigate the expression profile and biological function of lncRNAs in urothelial cancer stem cells and explore whether they can be biomarkers for prediction of clinical characteristics for bladder cancer patients.Methods Microarray analysis was performed to identify differentially expressed lncRNAs in cancer stem cells and parental cancer cells.Expression profiles were validated by Coding Potential Caculator analysis,real-time polymerase chain reaction.By performing in vitro sphere formation assays,J82 cells with lentivirus-based knockdown of lncRNA-BCSC (bladder cancer stem cells) were used to confirmed its function.Samples were obtained from patients who underwent TURBT between January 2009 and December 2012.All tissues were initially confirmed by pathologists.The association of the clinicpathologic of bladder cancer and lncRNA-BCSC expression was analyzed by x2 analysis.Results 750 lncRNAs were highly expressed from microarray analysis in bladder cancer stem cells.Among these,lncRNA-BCSC was identified as potentially maintaining self-renewal of cancer stem cells.Knockdown of this transcript in J82 cells inhibited spheroid formation.lncRNA-BCSC expression were higher in 54.8％ (57/104) bladder cancer tissues.Moreover,using x2 analysis,lncRNA-BCSC expression in primary tumors was found to be a predictor for recurrence following transurethral resection in patients with nonmuscle-invasive bladder cancer (P =0.009).Conclusions Our study provides strong evidence that lncRNA-BCSC are indispensable modulators of self-renewal ability of bladder cancer stem cells.Overexpression of lncRNA-BCSC may have a predictive value for early recurrence in patients suffering from nonmuscle-invasive bladder cancer.

Objective:To explore the inhibitory effect of dihydroartemisinin (DHA ) on the growth of neuroblastoma cells,and to clarify the anti-tumor mechanism of DHA.Methods:The experiment was divided into blank control group and DHA groups (the final concentrations of DHA were 0.05, 0.50, 5.00 and 50.00μmol·L-1 ).The proliferation rates of neuroblastoma SH-SY5Y cells after treated with DHA were examined by MTT assay;the changes of cell cycle of SH-SY5Y cells after treated with DHA were examined by flow cytometry;the expression levels of cyclin D1 and caspase-3 proteins were detected by ELISA and Western blotting methods.Results:The proliferation of SH-SY5Y cells 24,48,and 72 h after treated with different concentrations of DHA were inhibited.Compared with blank control group,the proliferation rates of SH-SY5Y cells in 0.50,5.00 and 50.00μmol·L-1 DHA groups were significantly decreased (P<0.05 or P<0.01).The density of cells was decreased with the increasing of DHA concentration.Compared with blank control group,the percentage of SH-SY5Y cells at SubG1 phase in 50.00μmol·L-1 DHA group was increased (P<0.05),and the percentage of cells at G0/G1 phase was increased first then was decreased;otherwise, the percentages of cells at S and G2/M phase were decreased.Compared with blank control group,the expression level of cyclin D1 protein in 50.00μmol·L-1 DHA group was decreased (P<0.05),but the expression level of caspase-3 protein in 50.00μmol· L-1 DHA group was increased (P<0.05).Conclusion:DHA could inhibit the proliferation through arresting the cell cycle and inducing the apoptosis of neuroblastoma cells.

Objective To study the feasibility and safety of performing nephrectomy together with the removal of complicated inferior vena cava tumor thrombus under profound hypothermia and arrested circulation. Methods After made the median thoraco-abdominal incision, the exploration of the abdominal organs was done. The right kidney, inferior vena cava and renal pedicle were well exposed then. After the whole body heparinization, cannulas were put into ascending aorta, superior vena cava, aortic root and right superior pulmonary vein. The body temperature was reduced to 20℃ with cardiopulmonary bypass unit and the extracorporeal circulation was stopped then. Cut open the inferior vena cava at vena renalis dextra ingress and the F16 urinary catheter was inserted into atrum dextra through inferior vena cava and inflated. The tumor thrombus was pulled out and the right kidney was removed. The inferior vena cava incision was sutured to close and the extracorporeal circulation was resumed and patient was re-warmed.Results The operation time was 330 min and the extracorporeal circulation time was 90 min, while the profound hypothermia with circulatory arrest time was 20 min. The estimated blood loss during operation was 400 ml and 6 unit red cells and 600 ml blood plasm were transfused. The patient was awaked 2.5 h after the operation, food intake resumed 4 days after operation and the patient was discharged on day 10 post-operatively. After 6 months'follow-up, there were no local recurrence and metastasis occurred. Conclusion The technique of profound hypothermia and circulation arrest could improve the safety and efficacy in the treatment of renal cell carcinoma with suprahepatic (level Ⅲ) caval tumor thrombus.

OBJECTIVETo observe the clinical efficacy differences between needle-knife combined with pseudo-ginseng-cake moxibustion and oral administration of western medication for primary knee osseous arthritis (PKOA) of blood stasis syndrome.

METHODSSixty cases of PKOA of blood stasis syndrome were randomly assigned to an observation group and a control group. Patients in the observation group (30 patients, 38 knees) were treated with needle-knife combined with pseudo-ginseng-cake moxibustion, while patients in the control group (30 patients, 36 knees) were oral administration of western medication. The treatment was given three weeks continuously. The knee osteoarthritis outcome score (KOOS) was adopted to observe the knee pain, symptoms and stiffness, daily life, sport and entertainment function, daily life quality score and total score in the two groups before and after treatment. The follow-up visit was performed two months after the treatment to evaluate the long-term efficacy.

RESULTSThe total effective rate was 97.4% (37/38) in the observation group, which was significantly superior to 77.8% (28/36) in the control group (<0.05). Each item of KOOS was increased in the observation group after treatment and follow-up visit (<0.05,<0.01); the KOSS in the control group was also increased (all<0.01) after treatment, and daily life and daily life quality score of knee in the follow-up visit were significantly increased (both<0.01). The scores other than sport and entertainment function score in the observation were significantly higher than those in the control group (all<0.01).

CONCLUSIONSThe needle-knife combined with pseudo-ginseng-cake moxibustion could significantly improve the symptoms of PKOA of blood stasis syndrome, which was superior to oral administration of western medication alone and had better long-term effects.

Objective: To investigate whether CD137-CD137L signaling could affect the secretion of mouse vascular smooth muscle cells (VSMCs) -derived exosomes through autophagy mediated Rab7 pathway. Methods: Primary thoracic aorta VSMCs from C57BL/6J mouse were obtained by tissue block adherence method. VSMCs between the third to fifth passages were used and VSMCs were divided into 4 groups: control group, CD137 agonist group, lentivirus control group, Rab7 lentiviral interference group. VSMCs in CD137 agonist group were treated with recombinant protein of CD137L (10 μg/ml), VSMCs in lentivirus control group were treated with lentiviral followed by recombinant protein of CD137L (10 μg/ml), VSMCs in Rab7 lentiviral interference group were treated with Rab7 lentiviral intervention followed by recombinant protein of CD137L (10 μg/ml). Western blot was used to detect the protein expression of LC3Ⅱ, p62, Rab7, CD9, CD81 and Hsc70. Fluorescence microscopy was used to track the changes of autophagy in cells infected with mRFP-GFP-LC3. Transmission electron microscope was used to observe the morphology and size of VSMCs-derived exosomes. The nanoparticle tracking analysis(NTA) was used to detect the concentration and size of exosomes in each group. Results: (1) The expressions of Rab7, LC3Ⅱ and p62 protein in VSMCs of CD137 activation group were significantly higher than those in control group (all P<0.05). The expressions of Rab7, LC3Ⅱ and p62 protein in Rab7 lentivirus interference group was lower than in CD137 activation group (all P<0.05), while the expressions were similar between the lentivirus control group and the CD137 activation group (all P>0.05). (2) The total number of fluorescent spots and yellow fluorescent spots in the VSMCs of the CD137 activation group were higher than those in the control group (all P<0.05), and the number of yellow fluorescent spots was higher than that of the red fluorescent spots in the VSMCs of the CD137 activation group ((50.3±0.9) vs. (10.3±1.5)/cell). The total numbers of fluorescent spots and yellow fluorescent spots in VSMCs of Rab7 lentivirus interference group were lower than those of CD137 activation group (both P<0.05), and the number of red fluorescent spots in VSMCs was higher than that of yellow fluorescent spots ((40.7±4.0) and (10.7±1.2)/cell) in the Rab7 lentiviral interference group. The total numbers of fluorescent spots and yellow fluorescent spots in the VSMCs were similar between the lentivirus control group and the CD137 activation group (all P>0.05). (3) Under transmission electron microscopy, the size of the VSMCs-derived exosomes was about 30-150 nm. The exosome markers (CD9, CD81) could be detected in vesicles by Western blot. NTA results showed that the concentration of VSMCs-derived exosomes was significantly higher in the CD137-activated group than in the control group (P<0.05), which was significantly lower in the Rab7 lentiviral interference group than in the CD137-activation group (P<0.05) and was similar between the lentivirus control group and the CD137 activation group (P>0.05). The expression of Hsc70 protein in exosomes secreted by CD137 activation group was higher than that in the control group (P<0.05). The expression of Hsc70 protein in exosomes was lower in Rab7 lentivirus interference group than in the CD137 activation group (P<0.05), which was similar between the lentivirus control group and the CD137 activation group (P>0.05). The expression of LC3Ⅱ protein in exosome was higher in CD137 activation group than in control group (P<0.05), which was lower in Rab7 lentivirus interference group than in CD137 activation group (P<0.05), which was similar between the lentivirus control group and the CD137 activation group (P>0.05). Conclusion The CD137-CD137L signaling may affect the secretion of mouse VSMCs-derived exosomes through modulating the Rab7 pathway mediated autophagy.

Objective To investigate the capsular polysaccharide (CPS) serotypes and molecular characteristics of carbapenem resistant hypermucoviscous Klebsiella pneumoniae (CR-HMKP) and study the possible mechanism of carbapenem resistance. Methods A retrospective study was conducted on 18 nonduplicate CR-HMKP strains which were collected from the First Affiliated Hospital of Nanchang University from 2012 to 2016. The clinical data were retrieved from medical records. The capsular serotypes, resistance genes and virulence factors were detected by polymerase chain reaction and DNA sequencing. Antimicrobial susceptibility testing was determined on VITEK 2 compact system. The CR-HMKP strains were characterized molecularly by using PCR, multilocus sequence typing (MLST) and pulsed field gel electrophoresis (PFGE). Modified carbapenem inactivation method was used to screen carbapenemase-producing strains. Plasmid conjugation transfer experiments were carried out to study transmission of carbapenem resistance. Results Eighteen (2.7％) CR-HMKP isolates were identified, which belonged to 4 serotypes, including wzi128-K1 (n=1), wzi206-K57 (n=1), wzi2-K2 (n=2), and wzi64-K14.64 (n=14). PCR and sequencing analysis identified blaNDM-1 gene in 2 CR-HMKP strains, blaKPC-2 gene in 17 strains, qnrS1 gene in 18 strains, blaCTX-M-3 gene in 3 strains, blaCTX-M-14 gene in 18 strains, blaTEM-1 gene in 16 strains, blaSHV-12 gene in 17 strains, and rmtB in 5 strains. All the 18 CR-HMKP strains carried virulence-associated genes, including rmpA (88.9％, 16/18), magA (5.6％, 1/18), iroN (83.3％, 15/18), aerobactin (27.8％, 5/18), rmpA2 (66.7％, 12/18) and mrkD (100％, 18/18). Three sequence types (STs) were identified by MLST, including ST11 (15 strains), ST86 (2 strains), and ST412 (1 strain). PFGE resulted in three major PFGE clusters, of which cluster A corresponds to ST1 isolates, and cluster B corresponds to ST86 isolates, and cluster C corresponds to ST412 isolates. All the blaKPC-2- positive strains belonged to ST11. Plasmid conjugation was successful in 5 (27.8％) of the 18 CR-HMKP isolates. Conclusions wzi64-K14.64 is the predominant capsule serotype of the CR-HMKP strains in this hospital. KPC-2 gene conjugationmay contribute to the emergence of CR-HMKP isolates. In addition, CRHMKP strain may be the highly prevalent ST11, and highly virulent CPS serotypes harboring K1/K2.

Objective:To investigate the expression levels of interferon-α receptor (IFNAR), interferon-stimulated gene factor 3(ISGF3), double-stranded RNA-activated protein kinase(PKR) and ribonuclease L (RNase L) in patients with chronic hepatitis B (CHB) treated with interferon.Methods:From July 2014 to June 2017, 41 treatment naive CHB patients were enrolled in the Department of Infectious Diseases, First Affiliated Hospital of Nanchang University. Eighteen patients were treated with polyethylene glycol interferon α-2b, and 23 patients were treated with conventional interferon. The mRNA and protein expression levels of IFNAR1, IFNAR2, ISGF3, PKR and RNase L in peripheral blood mononuclear cells (PBMC) were detected by reverse transcription polymerase chain reaction and Western blot, respectively. The differences of these molecular expression levels in PBMC between the effective and ineffective groups were compared. The data were analyzed by t test. Results:After 24 weeks of treatment, 25 cases were effective, while 16 cases were ineffective. At four weeks of treatment, the mRNA expression levels of IFNAR1, IFNAR2 and PKR in PBMC of the effective group were 0.748±0.129, 1.169±0.125 and 1.047±0.091, respectively, which were all higher than those in the ineffective group (0.591±0.021, 0.689±0.059 and 0.791±0.033, respectively). The differences were statistically significant ( t=-4.304, 16.482 and -5.346, respectively, all P<0.01). The mRNA expressions of ISGF3 and RNase L in PBMC of the effective group were 0.739±0.159 and 0.780±0.140, respectively, while those in the ineffective group were 0.690±0.035 and 0.733±0.122, respectively, which were not significantly different ( t=-0.160 and -1.443, respectively, both P>0.05). The mRNA expression levels of IFNAR1, IFNAR2, ISGF3, PKR and RNase L at baseline, week eight, 12 and 24 of treatment in the effective group were all higher than those in the ineffective group (all P<0.01). The protein expression levels of IFNAR1, IFNAR2, ISGF3, PKR and RNase L in the effective group were all higher than those in the ineffective group (all P<0.01). Conclusion:After interferon treatment, the mRNA and protein expression levels of IFNAR1, IFNAR2, ISGF3, PKR and RNase L in PBMC of CHB patients are all increased, especially IFNAR2 and PKR levels increase in the early stage of treatment (four weeks).