OBJECTIVE To explore urinary tract infection.METHODS Totally 750 clinical isolates of urinary tract infection were collected from patients who were cured in our hospital from 2001 to 2003,then analyzed the kinds of these bacterials and sensitive rate to antibiotics.RESULTS The percentage of Gram-negative bacilli was 53.60%,in which Escherichia coli was 38.13%;The percentage of Gram-positive cocci was 35.20%,in which Enterococcus were 16.53%;the percentage of fungi was 11.20%.The resistance rate of Escherichia coli to ampicillin,amoxicillin/clavulanic acid,quinolones,nalidixic acid and SXT was 80.54%,57.69%,45.00-71.00%, 71.23% and 65.14%,respectively,and the resistance rate to amikacin was lower than to gentamicin(5.82% vs 39.11%,P

In order to derive dynamic pulse rate variability (DPRV) signal from dynamic pulse signal in real time, a method for extracting DPRV signal was proposed and a portable mobile monitoring system was designed. The system consists of a front end for collecting and wireless sending pulse signal and a mobile terminal. The proposed method is employed to extract DPRV from dynamic pulse signal in mobile terminal, and the DPRV signal is analyzed both in the time domain and the frequency domain and also with non-linear method in real time. The results show that the proposed method can accurately derive DPRV signal in real time, the system can be used for processing and analyzing DPRV signal in real time.
Electrocardiography ; Heart Rate ; Monitoring, Physiologic ; Signal Processing, Computer-Assisted

Objective To construct a lentiviral vector carrying CRKL gene RNA interfering( RNAi ).Methods The CRKL RNAi was selected and subcloned into the lentiviral vector,pGCL-GFP(including U6 promotor and green fluorescent protein),generating the lentiviral vector LV-shCRKL.The corrected CRKL was confirmed by endoenzyme digestion ,sequencing.Recombinant lentiviruses were produced by 293T cells following the eo-transfection of LV-shCRKL,with the packaging plasmids pHelper1.0 and pHelper2.0.The virus titer was detected by GFP expressions in 293T cells.Results Plasmid LV-shCRKL carried the correct sequence.The recombinant lentiviruse LV-shCRKL could be produced by co-transfection of LV-shCRKL to 293T cells.Conclusion The recombinant lentiviruse vector LV-shCRKL is constructed successful.

Objective To investigate the role of nuclear factor(erythroid?derived 2)?like 2(Nrf2)in load?driven bone metabolism in Nrf2 knock?out(KO)mice. Methods The hybridized mice in the same brood were selected through PCR detection and were divided into two groups,i.e.,the Nrf2 knockout(KO)group and the wild?type(WT)group. Ulna of mice was loaded with 4 000 peak microstrains at 2 Hz for 3 consecutive days (120 cycles/day)as scheduled,the relative mineralizing surface(rMS/BS)and the relative bone formation rate(rBFR/BS)of ulna were measured for the two groups. Results Load?induced bone formation was suppressed in KO mice. Compared to the WT control,the relative bone formation rate was roughly 84%lower in KO mice(P<0.01). Conclusion The loss?of?function mutation of Nrf2 in bone diminishes load?driven bone formation.

Objective To detect the mRNA and protein expression of downstream quinine nicotinamide adenine dinucleotide (NADH) dehydrogenase 1 (NQO1) and heme oxygenase 1 (HO-1) induced by blood nuclearrelated factor E2 (Nrf2) in the peripheral blood of those exposed to arsenic in the endemic area of coal arsenic poisoning in Guizhou Province,and to discuss its role in the process of occurrence and development of liver injury due to coal arsenic poisoning.Methods Jiaole and Changqing villages in coal-burning-borne arsenism areas in Xingren County of Guizhou were selected as the survey sites,and 161 cases of arsenic-exposed residents were selected as the arsenic exposed group based on physical examination.They were divided into non-patient group (21 cases) and patient group (140 cases) according to the Diagnostic Criteria of Endemic Arsenism (WS/T 211-2001),and the patient group was further divided into mild hepatosis group (52 cases),moderately severe hepatosis group (36 cases) and non-apparent hepatosis group (52 cases) according to the Diagnostic Criteria of Occupational Chronic (GBZ 59-2010).Moreover,45 residents from one village neighboring to non-epidemic area were selected as controls.Peripheral blood samples were collected from all subjects.And mRNA expression of NQO1 and HO-1 were detected by RT-qPCR.Content of NQQ1 and HO-1 were detected by enzyme linked immunosorbent assay (ELISA).Results (①)Results of mRNA expression of NQ01 and HO-1:the relative expression level of mRNA of NQO1 and HO-1 in peripheral blood went up gradually as the degree of liver injury of population exposed to arsenic increased (F =5.548,10.961,all P ＜ 0.05);thereinto,the relative expression level of mRNA of NQO1 of mild hepatosis group (median:0.918) was higher than that of the control group (0.576,P ＜ 0.05),the relative expression level of mRNA of NQO1 of moderately severe hepatosis group (1.243) was higher than those of control group,non-patient group (0.653),non-apparent hepatosis group (0.636) and mild hepatosis group (all P ＜ 0.05);the relative expression level of mRNA of HO-1 of non-patient group (1.059),non-apparent hepatosis group (1.225),mild hepatosis group (1.553) and moderately severe hepatosis group (1.604) were higher than that of control group (0.767,all P ＜ 0.05);the relative expression level of mRNA of HO-1 of mild hepatosis group was higher than that of non-patient group (P ＜ 0.05);the relative expression level of mRNA of HO-1 of moderately severe hepatosis group was higher than those of non-patient group and non-apparent hepatosis group (all P ＜ 0.05).②)Results of protein expression of NQO-1 and HO-1:the level of protein expression of NQO1 and HO-1 in serum went up gradually as the degree of liver injury of population exposed to arsenic increased (F =19.685,17.725,all P ＜ 0.05).Thereinto,the protein expression of NQO1 and HO-1 of non-apparent hepatosis group,mild hepatosis group and moderately severe hepatosis group [NQO1:(6.272 ± 0.744),(6.336 ± 0.628),(6.714 ± 0.540) U/L;HO-1:(45.150 ± 4.813),(45.283 ± 5.049),(48.610 ± 5.365) U/L] were higher than those of control group and non-patient group [NQO1:(5.550 ± 0.730),(5.750 ± 0.427) U/L;HO-1:(39.856 ± 5.249),(42.375 ± 3.014) U/L,all P ＜ 0.05],the protein expression of NQO1 and HO-1 of moderately severe hepatosis group were higher than those of non-apparent hepatosis group and mild hepatosis group (all P ＜ 0.05).Conclusion The expression of mRNA and protein of NQO1 and HO-1 is closely related to the occurrence and development of liver injury due to arsenic exposure in coal arsenic poisoning areas.

Objective To investigate the effect of sodium arsenite (NaAsO2) on the expression of miRNA-21 (miR-21) mediated by transcription factor ETS-1 in human normal hepatocytes (L-02).Methods Dose-effect study:The L-02 cells were treated with different doses of NaAsO2 [0.0 (control),2.5,5.0,10.0,20.0,40.0 μmol/L] for 24 h.Time-effect study:L-02 cells were exposed to 0 (control) and 20 μmol/L NaAsO2 for 12,24,36 and 48 h (n =6).ETS-1 and miR-21 were treated with ETS-1 shRNA and miR-21 inhibitor,respectively.The cells treated with ETS-1 shRNA (100 nmol/L) were divided into 4 groups:①ETS-1 shRNA NC treatment alone (control group);②ETS-1 shRNA NC combined with NaAsO2 (20 μ,mol/L) treatment group;③ETS-1 shRNA treatment alone group;④Treatment with ETS-1 shRNA and NaAsO2 (20 μmol/L) group.The MiR-21 inhibitor (100 nmol/L) treated cells were also divided into 4 groups:① miR-21 inhibitor NC treatment (control group);② miR-21 inhibitor NC combined with NaAsO2 (20 μmol/L);③miR-21 inhibitor group;④miR-21 inhibitor combined with NaAsO2 (20 μ mol/L) treatment group.The expression of ETS-1 mRNA and miR-21 were detected by quantitative real-time PCR (qRT-PCR);the protein expression of ETS-1 was detected by Western blotting.Results Dose-effect study:The expression of ETS-1 mRNA in the groups of 0.0 (control),2.5,5.0,10.0,20.0 and 40.0 μmol/L was 1.008 ± 0.028,1.552 ± 0.029,1.697 ± 0.050,1.842 ± 0.077,2.233 ± 0.096 and 2.235 ± 0.092;miR-21 expression was 1.025 ± 0.094,1.552 ± 0.072,1.683 ± 0.066,1.915 ± 0.171,2.337 ± 0.195 and 2.592 ± 0.177;the expression of ETS-1 protein was 1.060 ± 0.045,1.267 ± 0.160,1.386 ± 0.087,1.723 ± 0.196,2.208 ± 0.122 and 2.284 ± 0.224,respectively,and the differences were statistically significant (F =47.797,8.959,65.748,all P ＜ 0.05),the NaAsO2 dose groups were significantly higher than those of the control group (all P ＜ 0.05),and there was a dose-effect relationship.Time-effect study:The expression of ETS-1 mRNA in L-02 cells was 1.253 ± 0.175,1.623 ± 0.220,1.771 ± 0.324 and 1.913 ± 0.251,respectively at 12,24,36 and 48 h;the expression of miR-21 was 1.502 ± 0.111,1.716 ± 0.113,1.979 ± 0.186 and 2.452 ± 0.304;the expression of ETS-1 protein was 1.196 ± 0.105,1.502 ± 0.076,1.651 ± 0.074 and 1.839 ± 0.139,respectively,there were significant differences between the groups (F =14.936,39.180,39.441,all P ＜ 0.05).The expression of various time points of exposure to NaAsO2 was significantly higher than those in the control group (1.044 ± 0.115,1.044 ± 0.124,1.108 ± 0.088,1.053 ± 0.061;1.092 ± 0.061,1.096 ± 0.169,1.024 ± 0.111,1.057 ± 0.146;1.020 ± 0.017,1.049 ± 0.121,1.024 ± 0.089,1.031 ± 0.124,all P＜ 0.05),and there was a time-effect relationship.ETS-1 shRNA and miR-21 inhibitor treatment:compared with ETS-1 shRNA NC combined with NaAsO2 (20 μmol/L),ETS-1 shRNA and NaAsO2 (20 μmol/L) could significantly inhibit the expression of ETS-1 (0.912 ± 0.238 vs 1.641 ± 0.225,P ＜ 0.05),and down-regulated the expression of miR-21 (1.313 ± 0.334 vs 2.363 ± 0.252,P ＜ 0.05).There was no significant difference of ETS-1 mRNA expression between miR-21 inhibitor and NaAsO2 (20.μmol/L) group (1.580 ± 0.077 vs 1.576 ± 0.065,P ＞ 0.05) compared with miR-21 inhibitor NC and NaAsO2 (20 μmol/L).Conclusions The expression of ETS-1 and miR-21 in L-02 cells is significantly higher than those in control.The high expression of ETS-1 mediates NaAsO2-induced miR-21 overexpression,which may be an important molecular mechanism of arsenic-induced expression dysregulation of human hepatic miRNAs and liver damage.

BACKGROUND: The survival rate of articular chondrocytes is low after traditional cryopreservation,and great differences existed in chondrocytes from surface layer and deep layer,which easily result in graft degeneration and lead to surgery failure.OBJECTIVE: To establish rabbit allograft models of graded frozen articular cartilages with holes made before cryopreservation and to observe the effect of holed cryopreservation on the rabbit articular cartilages.METHODS: Osteochondral plugs taken aseptically from 2 months old rabbits were randomly divided into 3 groups: the experimental group,making holes(3 mm× 3 mm)in articular cartilages and graded freezing; non-hole graded freezing group,non-making holes and graded freezing; cryopreservation group: non-making holes and rapid freezing.The grafts were thawed and transplanted into the relevant articular cartilage defects of recipient rabbits.The grafts differences were observed by gross observation,histochemistry and immunohistochemistry staining.RESULTS AND CONCLUSION: The gross observation,histochemistry and immunohistochemistry staining of the experimental group were superior to the cryopreservation group.Though there were no significant differences between the non-hole graded freezing group and the experimental group,however,the experimental group enhanced the protective effect on cartilage tissue in the middle layer.The graded cryopreservation of articular cartilage gets an advantage over rapid cryopreservation.And the articular cartilage with holes could be preserved successfully in graded cryopreservation,which assures the survival and function of chondrocytes and slows down degrading process of the articular cartilage tissue after thawed and transplanted.

Objective To detect the combining capacity of peripheral blood NF-E2-related factor 2 (Nrf2)of arsenic-exposed residents in the coal-contaminated arsenism area in Guizhou with the sequence of downstream antioxidant response element (ARE) as well as antioxidase gene expression,and to provide a basis for in-depth revelation of arsenic oxidative damage mechanism to human body.Methods Jiaole and Changqing villages in coal-burning-borne arsenism areas in Xingren County of Guizhou were selected as the survey spots,and 161 cases of arsenic-exposed residents were selected as the arsenic exposed group on the basis of physical examination.They were divided into non-patient group (21 cases) and patient group (140 cases) according to the Diagnostic Criteria of Endemic Arsenism (WS/T 211-2001),and the patient group was further divided into mild hepatosis group (52 cases),moderately severe hepatosis group (36 cases) and non-apparent hepatosis group (52 cases) according to the Diagnostic Criteria of Occupational Chronic (GBZ 59-2010).Moreover,45 residents from one village neighboring to non-epidemic area were selected as controls.The hemocyte nucleoprotein was extracted from peripheral blood in the sampling subjects.The combining capacity of peripheral blood Nrf2-ARE was tested by electrophoretic mobility shift assay (EMSA),and the relative expression quantity of Cu/Zn superoxide dismutase (Cu/Zn-SOD) and glutathione peroxidase 1 (GSH-Pxl) mRNA was tested with real-time fluorescence quantification PCR (qPCR).Results The testing results of Nrf2-ARE combining capacity showed that the difference of Nrf2-ARE combining capacity between groups was statistically significant (F =116.033,P ＜ 0.05).Compared with the control group (3.14 ± 1.34),the Nrf2-ARE combining capacity was higher in the non-apparent hepatosis group (5.17 ± 2.06),mild hepatosis group (13.13 ± 4.84) and moderately severe hepatosis group (32.35 ± 14.76,all P ＜ 0.05);compared with the non-patient group (5.15 ± 3.23) and non-apparent hepatosis group,the Nrf2-ARE combining capacity of mild hepatosis group and moderately severe hepatosis group was higher (all P ＜ 0.05);compared with mild hepatosis group,the Nrf2-ARE combining capacity of moderately severe hepatosis group was higher (P ＜ 0.05).The results of Cu/Zn-SOD and GSH-Pxl mRNA expression showed the relative expression quantities of Cu/Zn-SOD [Median (M):1.127 8,1.257 8,1.632 0] and GSH-Pxl (M:1.334 5,1.940 9,2.062 6) mRNA of non-apparent hepatosis,mild hepatosis,moderately severe hepatosis groups were higher than those of the control groups (M:0.961 8,0.884 3),respectively,and their differences were statistically significant (x2 =13.065,19.934,all P ＜ 0.05).The relative expression quantities of GSH-Pxl mRNA of mild hepatosis group and moderately severe hepatosis group were higher than that of the non-patient group (M:1.248 4),and their differences were statistically significant (all P ＜ 0.05).The correlation analysis indicated the Nrf2-ARE combining capacity was positively related to the Cu/Zn-SOD and GSH-Px1 mRNA expression (r =0.271,0.292,all P ＜ 0.01).Conclusion The increase of Nrtf2-ARE combining capacity by arsenic participates is involved in the regulation of Nrf2 downstream antioxidase gene expression,and this process possibly participates in occurrence and development of coal-burning-borne arsenism and hepatic injury.

Objective To investigate the change in expressions of lipoxin A4 (LXA4) and lipoxin A4 receptor (ALX) in rats with fat embolism syndrome(FES).Metbods Sixty healthy male SD rats were assigned to control group and FES group which was subgrouped at l,6,12 and 24 h according to the random number table,with 12 rats each.Allogeneic perinephric fat (0.706 ml/kg) was injected to rat caudal veins in FES group.Instead isotonic saline in an equal volume was given to rats in control group.Lung samples were harvested from each group to detect pathological morphology,concentration of total protein and LXA4 in bronchoalveolar lavage fluid (BALF),lung weight to dry ratio (W/D),and activity of myeloperoxidase (MPO) and ALX mRNA.Additional 40 SD rats were divided into control group,FES 24-hour group,BML-1 11 + FES 24-hour group,and Boc-2 + FES 24-hour group according to the random number table,with 10 rats each.Pathology of lung tissue was observed using microscopy and expression of lung MPO mRNA was detected.Results Lung tissues in FES group were seriously injured compared with control group.Total protein concentration in BALF was (71.12 ± 11.05) μg/ml in FES 12-hour group,significantly increased compared to (29.82 ± 0.64) μg/ml in control group (P ＜ 0.05).LXA4 concentration in BALF was (2.72 ± 0.24) ng/ml in FES 24-hour group,significantly higher than (0.69 ±0.05)ng/ml in control group (P ＜ 0.05).Lung W/D value was 9.13 ±0.83 and 9.60 ±0.86 respectively in FES 6-hour and 12-hour groups,higher than 3.09 ±0.10 in control group (P ＜0.05).Activity of MPO in lung tissue was (0.74± 0.07)U/g and (0.53 ±0.08)U/g respectively in FES 6-hour and 12-hour groups,significantly higher than (0.19 ± 0.03) U/g in control group (P ＜ 0.05).Expression of ALX mRNA was 3.99 ± 1.09 in FES 24-hour group,significantly higher than 1.00 ±0.21 in control group (P ＜0.05).Expression of MPO mRNA was lower in BML-111 + FES 24-hour group (0.69-0.08) and was higher in Boc-2 + FES 24-hour group (2.05-0.14),when compared to 1.52 ±-0.07 in FES 24-hour group (P＜0.05).Conclusion LXA4 mainly involves in the resolution of inflammation in FES rats,which may be achieved at least in part by binding to ALX.

Metformin is a first-line drug for the treatment of type 2 diabetes mellitus. Recently, some clinical studies have been published reporting a reduced incidence of various neoplastic disease (eg. breast cancer, endometrial cancer and gastrointestinal cancer) in diabetic or nondiabetic patients treated with metformin. Metformin inhibits the mammalian target of rapamycin by activating liver kinase B1 (LKB1)/adenosine 5'-monophosphate-activated protein kinase (AMPK) or decreases blood insulin levels, resulting in inhibition of cancer cell growth. There are also many other mechanisms involved in anti-tumor effect of metformin. Although metformin has significant effect on cancers, the prospects for it as an alternative treatment modality and mechanism in various kinds of tumors need further research.