Objective To develop a dual real-time fluorescent RT-PCR method for rapid detection of enterovirus(EV)and en terovirus type 71(EV71).Methods Specific primers and probes were designed and the dual real-time fluorescent RT-PCR reaction system was established.The quantitative standard curve was drawn;its sensitivity and precision were evaluated.Feces and throat swab specimens of 109 clinical patients with hand foot and mouth disease were collected and tested by using this method.Then the obtained results were compared with those detected by commercial EV71 PCR kit.Results The relative coefficient(2)of EV and EV71 standard curve established by the dual real-time fluorescent RT-PCR method were both 0.998.Its sensitivity reached 0.5 TCID50/mL for detecting EV and 0.05 TCID50/mL for detecting EV71.The within-run precision for detecting EV and EV71 was ＜3％ and total precision≤4％.The results showed good specificity for the detection of enterovirus and non-enterovirus.In 109 detected clinical samples,84 cases of EV positive samples were detected,in which 56 cases were EV71 positive with the total positive rate of 51.4 ％,which was consistent with the result of simple fluorescent RT-PCR commercialization kit(P=1.000).Conclusion The established dual real-time fluorescent RT-PCR method has high sensitivity and good stability,which has an important significance for early high throughput rapid diagnosis of hand foot and mouth disease.

Objective To improve the effects of health education(HE)and satisfaction degree on HE of inpatients by executing hierarchical full-responsibility nursing. Methods Fourteen wards were randomly chosen form the hospital and divided into the control group and the experimental group, each group having 7 wards. The control group carried out routine holistic nursing model, and the experimental group carried out a new hierarchical full-responsibility nursing system. Reforming nursing scheduling and diminishing the nursing unit to assure that the patients acquired the continous and stable nursing service when they were in hospital. The effect and the degree of satisfaction of HE were compared between two groups after 6 months. Results The effects of HE in the experimental group were significantly better than those in the control group, and the degree of satisfaction on HE in the experimental group was much higher than in the control group. Conclusions The hierarchical full-responsibility nursing system can give patients systematic and normalized HE. It can significantly improve the effects and the degree of satisfaction on HE.

Objective To compare the potential cytotoxicity induced by Veratrum nigrum coadministered with Panax ginseng, and to provide experimental evidence on the mode of herb-herb interaction based on human liver drug metabolizing enzymes.Methods The effect of V.nigrum and coadministration on cultured human hepatoma (HepG2) cells was investi-gated by detecting morphological changes , cell viability , cytomembrane integrity and apoptosis after the cells were treated for 24 h.The mRNA expression levels of drug metabolizing enzymes influenced by P.ginseng were determined by real-time quantitative reverse-transcriptase polymerase chain reaction .Results V.nigrum coadministered with P.ginseng had a better inhibitive effect on the growth of HepG2 cells at the IC50value of (15.18 ±1.03) mg/ml than at the value of IC50 (21.46 ±1.10) mg/ml of V.nigrum.Coadministration more significantly raised the LDH level in cell culture medium than at the same dose of V.nigrum.Moreover, in coadministration group, compared with the same dose of V.nigrum,the total apoptosis and necrosis of HepG2 cells were significantly increased .P.ginseng had effect on the expression of CYP3A4, CYP1A1, CYP1A2, CYP2B6 and CYP2E1 mRNA.Conclution Compatibility of medicines in a prescription also has herb-herb interactions based on drug metabolizing enzymes .The interaction mode is that the P.ginseng inhibits and induces CYPs and the modulated CYP isozymes ,inturn,have an impact on the metabolism of constituens in coadministered herbs causing herb-herb interaction .

Objective To study the inhibitory effect of mycophenolate mofetil (MMF) on expression of TGF-? in pulmonary fibroblasts (WI26 cells) and its application value.Methods Pulmonary fibroblasts were divided into control group,MMF group,TGF-? group,and MMF+TGF-? group,after routine culture.Expression of TGF-?-induced COL1A1 was detected by reverse transcriptase-polymerase chain reaction (RT-PCR).Expression of COL1 proteins was detected by Western blot analysis and chloramphenicol acetyl transferase assay,respectively.Difference in contractility of collagen fibers and migration of cells was observed in collagen matrix contraction test and cell scratch test.Results The expression level of COL1A1 mRNA was higher in MMF group than in control group 24 and 48 h after treatment with MMF (77.0%?2.9% vs 38.0%?3.7%),and was lower in MMF group than in control group 24 and 48 h after treatment with TGF-? and MMF+ TGF-? (134.0%?3.1% vs 189.0%?2.4%,and 95.0%?2.7% vs 71.0%?3.3%,P

Objective To assess the impact factors of surgical site infection(SSI) in the department of general surgery,the improve the quality of the target of monitoring,provide clinical theoretical basis for reducing the incidence of SSI.Methods In 2015,920 patients who underwent general surgery was took in the targeted monitoring of SSI.SPSS19.0 software was used to analyzing the data.Results The infection rate was 4.35％;Surgical site infection rate was rising,with the increase of NNIS.17 pathogens were isolated,including 11 Escherichia colis which was the most.The incidence of the SSI was 2.40％ between two groups in the patients who underwent the elective surgeries 10.85％,in the patients who underwent emergency surgery.there was significant difference between two groups(x2 =27.997,P＜0.05).The type Ⅱ surgical incision was smain type in the department of general surgery,the incidence of the typeⅡ surgical incision was 2.27％,the incidence of the typeⅢ surgical incision was 21.90％,no SSI occurred in the type Ⅰ surgical incision;SSI incidence of surgery time which was more than 3 h was 7.27％,less than 3 h was 3.71 ％,there was significant difference between two groups(x2 =4.136,P＜0.05);the SSI incidence of the incision length ≥10 cm was 13.11 ％,less than 10 cm was 1.82％,the difference was statistically significant (x2=48.966,P＜0.05).Conclusion NNIS score,wound type,type of surgery,duration of surgery may become the risk factors SSI.

Objective To compare volumetric‐modulated arc therapy(VMAT) with intensity‐modulated radiation therapy (IMRT) for brain metastases with regard to the dosimetric character .Methods Sixty patients who were diagnosed with brain me‐tastases were included in this study .The target area received two dose levels using late addition amount technique ,WBRT (30 Gy/10 F) with following addition (20 Gy/10 F) to 59 Gy .For a fair comparison ,VMAT and IMRT treatment plans were respectively designed for every patient with the same dosimetric constraints .Dosimetric comparisons between VMAT and IMRT plans were ana‐lyzed to evaluate :target coverage and homogeneity ,conformity of PTV ;sparing of OARs ;monitor units (MUs) .Results Two treatment plans all reached the treatment need .When compared with IMRT ,there was no significant difference in Dmean of eyeball , len ,optic never ,visual chiasma ,parotid ,brain stem ,and external auditory canal of VMAT (P>0 .05) .The Dmax of eyeball ,len ,pa‐rotid ,and external auditory canal of VMAT were lower than that in IMRT group (P<0 .05) .The VMAT group has the less MUs (P=0 .017) and less treatment time .Conclusion VMAT can reach the big‐dose radiotherapy need on brain metastases clinically . There are no significant diffference between VMAT and IMRT on Dmax ,Dmean ,CI ,and HI .The Dmax of eyeball ,len ,parotid ,and external auditory canal of VMAT were lower than that in IMRT group .The VMAT can reduce the radiotherapy time .

Objective To establish the cell model of ethanol-induced liver injury and explore the protective effects of tea poly-phenols (TP)on ethanol-induced liver injury .Methods Cell morphology were observed by microscope ,and then alanine aminotrans-ferase (ALT) ,nmda transaminase (AST) ,gamma GGTP ,GGT and ROS changes were detected .Results Alcohol maked L02 hepa-tocyte fatty degeneration .Compared with ethanol group ,steatosis in TP + ethanol group was lighter ,its ALT ,AST ,GGT content and intracellular ROS reduced .Conclusion TP can decrease cell fatty change degree in vitro experiments ,improue the enzymology indexes ,reduce the generation of reactive oxygen species to avoid liver damage .

Objective To investigate the clinical significance of retinal binding protein (RBP) and Cystatin C in blood and urine in patients with chronic kidney disease (CKD).Methods One hundred and twenty-one patients with CKD in Nephrology Laboratory of The People's Hospital of Xinjiang Uygur Autonomous Region from Aug.2012 to Jan.2013 were served as the case group.Sixty healthy check-up people were considered as control group.Enzyme linked immunosorbent assay was used to detect RBP and Cystatin C in urine,and the immune turbidimetry testing was applied to detect RBP and Cystatin C in blood.The receiveroperating characteristic (ROC) curve and area under curve (AUC) were applied to evaluate the sensibility and diagnostic value of indexes in CKD.Results The positive rate of RBP,Cystatin C in urine and blood were 89.6％,92.6％,52.5％,59.5％ respectively of 12 patients in case group,higher than those in control group (all value were 0,P ＜0.001).ROC curve showed that the sensitivity of urine RBP and Cystatin C were higher than that in blood of case group.AUC of RBP,Cystatin C level in urine and blood were 0.915,0.974,.655,and 0.623 respectively.And 95％ confidence interval were 0.877-0.954,0.956-0.992,0.575-0.736,0.543-0.702 respectively,and the differences are statistically significant (P ＜ 0.001).Conclusion The sensitivity of urine cystatin C and RBP are significantly higher than that in blood,which might be served as a diagnostic marker of CKD and can provide important basis for clinical diagnosis.

OBJECTIVETo observe mainfestations of syndrome and biochemical indices of hypertensive model rats with excessive accumulation of phlegm-dampness syndrome (EAPDS), and to explore its possible pathological mechanism.

METHODSEAPDS rat model was prepared in 50 Wistar rats by feeding with high fat forage. Meanwhile, a normal control group consisting of 10 Wistar rats was set up by feeding with normal forage. After 25-week continuous feeding, 22 rats with body weight (BW) and blood pressure (BP) exceeding 25% those of the control group were selected as a model group. BW, BP, blood lipids, and related serological indicators were detected in all rats. Morphological changes of target organs were observed. mRNA expression levels of leptin receptor (LepR), Janus kinase2 (Jak2), signal transducer and activator of transcription 3 (Stat3), suppressor of cytokine signaling-3 (Socs3), angiotensin II receptor type 1 (AT1), angiotensin II receptor type 2 (AT2), phosphatidylinositol 3 kinase (P13K), serine threonine kinase (Akt), nuclear factor of kappa B (NF-κBp65), inhibitor of nuclear factor kappa-B kinase α (IKKα), NF-kappa-B inhibitor β (lKKβ), NF-kappa-B inhibitor α (IKBα), and AMP-activated protein kinase (AMPK) were detected by quantitative real-time PCR (qPCR). Expression levels of AT1 and LepR in aorta were detected by immunohistochemical assay and Western blot respectively.

RESULTSCompared with the control group, BW, BP, and blood lipids increased; serum levels of leptin (Lep) , Ang II, Hcy, ET-1, TNF-α, IL-6, and p2-MG increased, but NO decreased in the model group (P < 0.05, P < 0.01). Aortal endothelial injury and smooth muscle cell proliferation occurred in the model group, accompanied with heart and renal injury. Compared with the control group, mRNA expression levels of LepR, Jak2, Stat3, Socs3, AT1 , PI3K, Akt, NF-κB p65, IKKβ, IKBα, and AMPK in aorta were up-regulated significantly (P < 0.05), while the expression of IKKa decreased (P < 0.05). Immunohistochem- ical staining showed, brownish yellow deposit of AT1 and LepR was obviously increased, with more extensively positive distribution. Western blot results showed, as compared with the control group, protein expression levels of AT1 and LepR obviously increased in the model group (P < 0.05).

CONCLUSIONSModel rats exhibited typical syndromes of EAPDS. They put up weight with fat abdomen, gloomy hair, poor appetite, hypersomnia, lowered activities , reduced food intake, loose stool, dark red tongue, white tongue with white, thick, greasy fur. Lep could be taken as one of objective indicators for evaluating hypertension rat model with EAPDS.

Animals ; Aorta ; Cell Proliferation ; Disease Models, Animal ; Hypertension ; physiopathology ; I-kappa B Proteins ; Interleukin-6 ; Leptin ; blood ; NF-KappaB Inhibitor alpha ; NF-kappa B ; Phosphatidylinositol 3-Kinases ; Rats ; Rats, Wistar ; Suppressor of Cytokine Signaling Proteins ; Transcription Factor RelA ; Tumor Necrosis Factor-alpha

[Objective] To observe the effect of cyclosporine A (CSA) on the activity and expression of MMP-2 and MMP-9 in renal tubular epithelial cells. [Methods] NRK52E cells were cultured until its reached confluent. Then NRK52E cells were exposed to different concentration of CSA (0, 0.42, 0.84, 4.2, and 8.4 μmoL/L) for 48 h or 72 h respectively. MMP-2 and MMP-9 activities were detected by gelatin zymography. The expression of MMP-2 and MMP-9 mRNA were detected by RT-PCR. [Results] MMP-2 activity and mRNA levels were decreased in a dose dependent manner after exposed to different concentration of CSA for 48 h or 72 h in NRK52E cells. Compared with control (CSA 0 μmoL/L), CSA 0.42, 0.84, 4.2, 8.4 μmoL/L decreased the MMP-2 activities to 27%, 24%, 11%, and 9% respectively; The differences are significant, P<0.05. But the MMP-9 activity and mRNA levels were increased after exposed to CSA for 48 h or 72 h in NRK52E cells. Compared with control group, CSA 4.2 μmoL/L exposure increased MMP-9 activity to 438% in 48 h, and 237% in 72 h; the differences are significant as well, P<0.05. [Conclusion] A dose-dependent decrease in the expression and activity of MMP-2, and the up-regulation of the expression and activity of MMP-9 by CSA in renal epithelial cells may related to CSA associated tubulointerstitial damage.