Objective To clarify whether the NADPH oxidases (NOXs) family contributed to the reactive oxygen species (ROS) production and subsequent interstitial fibrosis in unilateral ureter obstruction (UUO) rats.Methods Male Wistar rats were randomly divided into sham operation group (n =8),sham operation + apocynin treatment group (n =8),UUO operation group (n =8) and UUO operation+apocynin treatment group (n =8).Either vehicle or apocynin (100 mg/kg per day) were given by gavage for 7 days after surgery.Rats were sacrificed at 7th day.ELISA was used to detect the activity of superoxide dismutase (SOD) and catalase (CAT),and the level of 8-iso-prostaglandin F2alpha (8-isoPGF2α) in renal tissue.Western blotting was used to detect the protein expressions of NADPH oxidase subunit NOX2 and NOX4,α-smooth muscle actin(α-SMA),collagen Ⅰ (COL-Ⅰ) and the level of ERK1/ 2 phosphorylation (p-ERK1/2).Results UUO rats with vehicle displayed increased oxidative stress,as measured by renal tissue 8-iso-PGF2α,accompanied with increased renal expression of NADPH oxidases (NOX2,1.5-fold and NOX4,1.7-fold,respectively),compared with sham-operated rats (P ＜0.05).Furthermore,vehicle treated UUO rats showed increased renal COL-Ⅰ and α-SMA levels,compared with sham-operated rats (P ＜ 0.05).ERK1/2 was also activated as detected by p-ERK1/2 expression in UUO rats with vehicle (P ＜ 0.05).Apocynin treatment significantly decreased renal tissue 8-iso-PGF2α level and expressions of NOX2 (-28.7％) and NOX4 (-31.0％) in UUO rats,respectively,compared with vehicle treated rats (P ＜ 0.05).And significant decrease of COL-Ⅰ (-26.4％) and α-SMA expression (-80.0％) were also observed (P ＜ 0.05).The activation of ERK1/2 in UUO rats was greatly inhibited by apocynin treatment (P ＜ 0.01).Despite the pronounced dysregulation of pro-oxidative NOXs family,no compensatory increase of antioxidative enzyme activities occurred.Conclusion The NOXs family contributes largely to the production of ROS and subsequent interstitial fibrosis after ureter ligation,and inhibition of the NOXs family may be a choice for preventing interstitial fibrosis.

Objective To investigate the influence of a combination of Chinese herbs,Astragalus membranaceus var.mongholicus and A ngelica sinensis (A&A)on the production and elimination of reactive oxygen species (ROS)and the underlying mechanism during the process of renal interstitial fibrosis in the obstructive kidneys of rats with unilateral ureteral obstruction(uuo).Method Male Wistar rats were randomly divided into control,sham,UUO and UAA (UUO+A&A)groups.The rats in UAA group were administered with A&A(14 g/kg)by oral gavage once daily:the ones in sham and UUO group were given with equal volumes of water.Three days after setting up models,pathological injury of renal tissue was evaluated.Level of total antioxidant capacity(T-AOC)and activity of CuZn superoxide dismutase(CuZn-SOD)in renal homogenates were measured bv spectrophotometry.Expression of NADPH oxidase subunits p47-phox,p22-phox and nintrotyrosine was analyzed by Western blot. Results Severe interstitial inflammatory cells infiltration, mild tubular atrophy and interstitial fibrosis were found in UUO group, which were alleviated by A&A administration. Compared to sham group, T-AOC of UUO group was not significantly changed, but the expression of the nitrotyrosine, NADPH oxidase subunits p47-phox and p22-phox was increased significantly (P<0.05). After A&A administration, T-AOC level was increased (2.5±1.1 vs 1.5±0.5, P<0.05) and the expression of nitrotyrosine was decreased (P< 0.05) in UAA group compared with UUO group. Additionally, the expression of p47-phox was reduced at day 3 (P<0.05), accompanied with a reduced expression of p22-phox (P< 0.05). CuZn-SOD activity was not significantly changed among the groups. Conclusion The inhibition of A&A in NADPH oxidase subunits p47-phox and p22-phox, which is responsible for reduction of oxidative stress, is associated with the alleviation of renal fibrosis in obstructive rat kidney.

Objective To evaluate a new computer-aided technique applicable for myocardial contrast echocardiography(MCE) to quantitate automatically calibrated myocardial contrast intensity(CD and to test the feasibility of calibrated CI in assessing myocardial perfusion. To analyze the relationship on myocardial perfusion,regional contractile function and cell apoptosis in stunned myocardium. Methods According to coronary occlusion and reperfusion at different times, rabbits were divided into three groups: 15 min occlusion/30min reperfusion (group Ⅰ ),30 min occlusion / 60min reperfusion (group Ⅱ ) and 120 min occlusion / 60min reperfusion (group Ⅲ ). MCE was performed on all rabbits at baseline,occlusion and after reperfusion,and its images were analyzed by a new computer-aided technique. Myocardial calibrated CI of each segment was measured automatically by software. Percentage wall thickening (WT) of each risk segment at each stage were also measured by echocardiography. The apoptotic index(AI) in regional left myocardial dysfunction was calculated by terminal deoxynucleotidyl transferease-mediated biotinylated deoxyuridine triphosphate nick end labeling(TUNEL ). Results (1) During occlusion, WT in the areas at risk decreased to zero or negative and the calibrated CI values were significantly lower than those at baseline ( P ＜0.01 ). After reperfusion, WT in all risk segment remained depressed, but calibrated CI significantly improved in group Ⅰ and Ⅱ while those remained unchanged in group Ⅲ. (2)AI in risk myocytes were (13. 70 ± 5.48 ) %, (36.25 ± 5.55 ) % and ( 62.06 ± 6. 70 ) %, respectively, both statistically significant difference between the two groups ( P ＜0.05 or P ＜ 0.01 ). AI were negatively correlated to WT and calibrated CI ( r = - 0. 87 and r = - 0. 77, P ＜0.05). Conclusions MCE with computer-aided technique can assess quantitatively myocardial perfusion and regional contractile function. Short-term ischemiareperfusion does not cause myocardial necrosis, but it will lead to myocardial cell apoptosis and the phenomenon of myocardial stunning. Prolonged ischemia, even if given sufficient reperfusion, can lead to apoptosis and necrosis simultaneously.

BACKGROUND:Stem cel therapy has been used for prevention and treatment of degenerative disc disease. Considering the special microenvironment in the intervertebral disc, the survival rate and differentiation ability of transplanted cels are decreased, which may lead to the poor efficacy of stem cel therapy. How to improve the survival ability and therapeutic effect of the transplanted cels is the focus of stem cel therapy for degenerative disc disease.
OBJECTIVE: To investigate the effects of cobalt chloride combined with hypertonic solution pretreatment on bone marrow mesenchymal stem cels that wil be transplanted for treatment of degenerative disc disease.
METHODS:(1)In vitro cel experiment: bone marrow mesenchymal stem cels were divided into three groups and subjected to normal culture medium (normal control group), 1% hypertonic mother solution (hypertonic group), 100 μmol/L cobalt chloride (hypoxia group), or 1% hypertonic mother solution plus 100 μmol/L cobalt chloride (combined group) for 1 week. Then, 2% hypertonic solution and 200 μmol/L cobalt chloride cobalt chloride were used to simulate the anaerobic and hypertonic environment intervenes in pretreated and untreated bone marrow mesenchymal stem cels for 24 hours. After that, RT-PCR was used to detect the expression of caspase-3 for apoptosis evaluation. (2)In vivo animal experiment: Sprague-Dawley rats were divided into model, cel transplantation and hypertonic plus hypoxic groups. Rat models of intervertebral disc degeneration were made in these three groups. After modeling, rats in these three groups were given no treatment, bone marrow mesenchymal stem cel transplantation or transplantation of bone marrow mesenchymal stem cels which were subjected to hypertonic and hypoxia pretreatments into the intervertebral disc. Two weeks later, immunohistochemistry and RT-PCR methods were used to detect cel distribution and related gene expression, respectively, thereby to evaluate the therapeutic effect of stem cels.
RESULTS AND CONCLUSION: (1)In vitro cel experiment: caspase-3 mRNA expression was significantly reduced in pretreated bone marrow mesenchymal stem cels compared with the untreated cels (P < 0.05). (2)In vivo animal experiment: compared with the control group, the caspase-3 and interleukin-1β in the intervertebral disc and a number of degenerative indexes were decreased in the cel transplantation. Compared with the cel transplantation group, these indicators had better outcomes in the hypertonic plus hypoxic group (P < 0.05). These findings indicate that bone marrow mesenchymal stem cels have therapeutic potential for degenerative disc disease, and have better adaptability and transplantation effects by hypertonic and hypoxia pretreatments.

Background Chronic low-level exposure to lead can damage the central nervous system and cause anxiety-like behavior. However, whether changes of blood metabolites occur in this process and its relationship with lead-induced neurobehavioral disorder remain unclear. Objective To explore the effects of chronic lead acetate (PbAc) exposure at different concentrations on anxiety-like behavior and serum metabolites and their relationships in mice, as well as the mechanism of lead exposure on neurobehavioral injury in mice from the perspective of metabolomics. Methods A total of 64 healthy 4-week-old C57BL/6J mice, half male and half female, were randomly divided into four groups: control group (normal drinking water), 20 mg·L⁻¹ PbAc group, 100 mg·L⁻¹ PbAc group, and 500 mg·L⁻¹ PbAc group. After 10 weeks of free drinking of water containing designed concentrations of PbAc, the mice were tested for anxiety-like behavioral changes by open field experiment. After the mice were anesthetized, blood was collected from the eyes, the serum was separated, and the effects of designed doses of lead exposure on metabolites in the serum of mice were compared by liquid chromatography with tandem mass spectrometry combined with principal component analysis and partial least squares discrimination analysis. Results The results of the open field experiment showed that the reductions in movement time spent in central area in the 100 mg·L⁻¹ and 500 mg·L⁻¹ PbAc groups compared with the control group were of statistical significance (P<0.05); the reduction in crossing times of central region in the 500 mg·L⁻¹ PbAc group was statistically significant compared with the control group (P<0.05); the increases in defecation frequency in the 100 mg·L⁻¹ and 500 mg·L⁻¹ PbAc groups were statistically significant compared to the control group (P<0.05). In both positive and negative ion modes, compared with the control group, 157 differential metabolites were screened out in the 20 mg·L⁻¹ PbAc group, of which 80 were up-regulated and 77 were down-regulated; 172 differential metabolites were screened out in the 100 mg·L⁻¹ PbAc group, of which 57 were up-regulated and 115 were down-regulated; 119 differential metabolites were screened out in the 500 mg·L⁻¹ PbAc group, of which 42 were up-regulated and 77 were down-regulated. The results of the KEGG enrichment analysis on the differential metabolites revealed alterations in metabolic pathways mainly involving primary bile acid biosynthesis, bile secretion, and cholesterol metabolism. Among the differential metabolites, norethisterone was positively correlated with the number of central region crossings (r=0.406, P<0.05); dihydrothymine was negatively correlated with the number of central region crossings (r=−0.346, P<0.05); lysophosphatidylcholine 22∶1 and lysophospholipid 14∶0 were negatively correlated with time spent in central region (r=−0.429, P<0.05; r=−0.374, P<0.05). Conclusion Chronic lead exposure induces anxiety-like behavior in mice, and this altered behavior is associated with altered metabolites in serum, with differential metabolites enriched primarily in the metabolic pathways of primary bile acid biosynthesis, bile acid secretion, and cholesterol metabolism.

OBJECTIVEThe imbalance between extracellular matrix (ECM) synthesis and degradation induces the excessive ECM deposition and thus renal fibrosis. The decoction (A&A) which is a combination of two Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, has been shown to alleviate ECM production in animal models of chronic kidney diseases. In this paper, the effect of A&A on ECM degradation was investigated with interstitial fibrosis in rats.

METHODMale Wistar rats were randomly divided into sham, unilateral ureteral obstruction (UUO) and UAA (UUO plus A&A administration) groups. After administration of A&A (14 g x kg(-1) x d(-1)) by gavage for 3, 7 and 10 days, morphological changes were evaluated by HE, PAS and Sirius red staining technique. The expression of plasminogen activator inhibitor-1 (PAI-1) and tissue-type plasminogen activator (t-PA), the activity of PAI-1 and t-PA were determined by ELISA. The activity of matrix metalloproteinases (MMP-9, MMP-2), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) were evaluated by gelatin zymography or reverse gelatin zymography, respectively.

RESULTMorphological analysis showed severe interstitial mononuclear cells infiltration, tubular atrophy, renal fibrosis and collagen expression in kidneys of UUO group, which was reduced by A&A administration (P < 0.05, UAA vs UUO group). Compared with the sham group, the expression of PAI-1 was significantly increased in UUO group by 63%, 91% and 112% at day 3, 7 and 10 respectively; and there were a remarkable decrease in UAA group by 44%, 43% and 52% at day 3, 7 and 10. The expression of active PAI-1 was strikingly increased in UUO group at day 3 [(30.5 +/- 23.8) ng x g(-1) vs. (0.0 +/- 0.0) ng x g(-1), P < 0.05)], day 7 [(36.5 +/- 11.2) ng x g(-1) vs. (0.0 +/- 0.0) ng x g(-1), P < 0.05)], and day 10 [(54.5 +/- 14.2) ng x g(-1) vs. (0.5 +/- 0.5) ng x g(-1), P < 0.05)]. The active PAI-1 was decreased in UAA group at day 7 [(14.9 +/- 0.5) ng x g(-1) vs. (36.5 +/- 11.2) ng x g(-1), P < 0.05] and day 10 [(15.4 +/- 4.8) ng x g(-1) vs. (54.5 +/- 14.2) ng x g(-1), P < 0.05]. The expression of t-PA was increased in UUO group only at day 3 [(58.1 +/- 16.5) microg x g(-1) vs. (30.1 +/- 17.3) microg x g(-1)], P < 0.05), meanwhile decreased in UAA group [(26.3 +/- 8.7) microg x g(-1) vs. (58.1 +/- 16.5) microg x g(-1), P < 0.05)]. But the expression of active t-PA was shown no significantly difference among the three groups. For MMP-2 and MMP-9 activity, they were significantly higher compared with the sham group in UUO group, but no significantly change after A&A treatment. The TIMP-1 activity was significantly increased in UUO group by 28% and 63% at day 7 and 10 respectively, significantly decreased in UAA group by 40% and 39% at the same time point.

CONCLUSIONThe anti-fibrosis effects of A&A might be associated with modulating the imbalance of PAs/PAIs system as well as MMPs/TIMPs system, thereby alleviate ECM accumulation and interstitial fibrosis.

Angelica sinensis ; chemistry ; Animals ; Astragalus Plant ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Extracellular Matrix ; metabolism ; Fibrosis ; Humans ; Kidney ; enzymology ; metabolism ; pathology ; Kidney Diseases ; drug therapy ; enzymology ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Plasminogen Activator Inhibitor 1 ; metabolism ; Rats ; Rats, Wistar ; Tissue Plasminogen Activator ; metabolism

Aim To investigate the roles of chloride channels in the apoptosis and apoptotic volume de-crease (AVD)induced by adriamycin in nasopharyn-geal carcinoma CNE-2Z cells.Methods Apoptotic rates were detected by flow cytometry,and the volume changes were measured by the time-lapse live cell ima-ging technique.The patch clamp technique was used to record whole-cell chloride currents.Results Adria-mycin induced apoptosis of CNE-2Z cells.An early ap-optotic volume decrease was observed in the cell trea-ted with adriamycin.The cell volume was decreased by about 10% in 2 h.Adriamycin activated a chloride current which showed outward rectification.The chlo-ride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB ) could inhibit the adriamycin-in-duced chloride currents,apoptosis and prevent cell shrinkage.Conclusions Our findings suggest that ad-riamycin causes cell apoptosis by activation of chloride channels.Chloride channels may be involved in the apoptosis and apoptotic volume decrease induced by adriamycin in CNE-2Z cells.

Objective: To investigate factors associated with awareness of prevention and control of Norovirus outbreak among primary and middle school staffs in the Pearl River Delta region，to provide evidence for Norovirus outbreaks prevention and control. Methods: A total of 320 primary and middle school staffs from 48 schools in 3 cities in the pearl river delta region were selected by multistage quota random sampling method and were interviewed face to face with questionnaire about knowledge of Norovirus and prevention and control of Norovirus outbreaks. Results: The awareness rate of Norovirus knowledge and prevention and control of Norovirus outbreaks among primary middle school and the total school staffs in the pearl river delta region were 35.37%,57.63% and 48.42%,respectively. The results of univariate analysis showed that the awareness rate of prevention and control of Norovirus outbreaks was significantly affected by duties, the total awareness rate among teachers(OR =0.22，95%CI=0.09-0.55),cooks(OR=0.09，95%CI=0.04-0.23)and cleaners(OR=0.02，95%CI=0.00-0.07)were far lower than school doctors(P<0.05). Conclusion The awareness of prevention and control of Norovirus outbreak among primary and middle school staffs in the Pearl River Delta region was needed to be improved. Valid implements, such as operation procedure, to build up prevention and control capability should be carried out that focus on specific job responsibilities.

Objective To investigate the clinical and imaging features of the aquaporin-4 (AQP4)antibody-negative binocular optic neuritis and to analyze the predictive factors of visual function outcome.Methods Fifty-eight patients with AQP4-negative binocular optic neuritis were reviewed and followed up from January 2014 to December 2015.Patients at baseline and at the end of follow-up were evaluated for visual function and neurological examination.All patients underwent optic nerve and brain MRI, cerebrospinal fluid and routine laboratory tests.Results AQP4 antibody-negative binocular optic neuritis accounted for 9.4%(58/615)of the total optic neuritis in the same period.At baseline, 99 eyes (85.3%,99/116)had best corrected visual acuity<0.1.At the end of follow-up, 31 eyes(26.7%,31/116) had best corrected visual acuity < 0.1. There were 43 cases (74.1%, 43/58) with multi-segment involvement of optic nerve at the baseline.Baseline visual acuity(P=0.005), early treatment response (P=0.011), and segment numbers of optic nerve involvement(P=0.025)were independently associated with end-point outcome of visual function.Forty-nine patients(84.5%,49/58)showed monophasic course in (3.1 ± 0.9) years follow-up period, 7 cases (12.1%, 7/58) had recurrence, and 2 cases (3.4%, 2/58) converted to neuromyelitis optic spectrum disorder (NMOSD). Conclusions AQP4 antibody-negative binocular optic neuritis is common and the recovery of visual function is not satisfied. Baseline visual function and the length of optic lesion in MRI is related to the end-point prognosis. Most patients performs the single phase course during the follow-up period.

Objective To compare the accuracy of electroencephalography (EEG) grading scale with amplitude-integrated electroencephalography (aEEG) in predicting poor outcomes (3-month), who sustained coma after cardiopulmonary resuscitation (CPR) in adults. Methods A retrospective study was conducted. The patients with post-anoxic coma admitted to intensive care unit (ICU) of Tongren Hospital, Capital Medical University from March 2010 to June 2017 were enrolled. EEG was registered and recorded at least once within 7 days of coma after CPR, while not being subjected to therapeutic hypothermia. General data, Glasgow coma scale (GCS), EEG grading and aEEG model were collected. According to Glasgow prognosis score (GOS) of 3-month outcome, patients were divided into poor prognosis group (GOS 1-2) and good prognosis group (GOS 3-5), and the differences of related indexes between the two groups were compared. The predictive ability of aEEG model and EEG grading for brain function prognosis was evaluated by receiver operating characteristic (ROC) curve. Results Fifty-four patients were included, with 31 males and 23 females, and age of (53.9±19.3) years. Among the EEG Young grades, 17 cases (31.5%) were grade 1, 4 cases (7.4%) were grade 2-5, and 33 cases (61.1%) were grade 6. Among the aEEG model grades, 26 cases (48.1%) had slow wave pattern grade 1, 23 cases (42.6%) had suppressed mode grade 4, 4 cases (7.4%) had status epilepticus mode grade 2, and 1 case (1.9%) had burst suppression mode grade 3. Thirty-six patients had poor prognosis 3-month after onset, 26 of them died and 10 had persistent vegetative state. The prognosis was good in 18 cases, including 16 cases with severe neurological disability and 2 cases with moderate neurological disability. There was no significant difference in gender, age, anoxic time between two groups with different prognosis, while the degree of consciousness disorder in poor prognosis group was more severe than that in good prognosis group (GCS score: 4.1±1.7 vs. 5.0±2.1, P < 0.05). The consistency test showed that different physicians had good consistency in EEG grading and aEEG model (Kappa values were 0.917 and 0.932, respectively). It was shown by ROC curve analysis that the area under ROC curve (AUC) of aEEG model and EEG grading for predicting poor prognosis of coma patients after CPR were 0.815 and 0.720, respectively (both P < 0.01); when the cut-off value of aEEG was 2.5, the sensitivity was 79.3%, the specificity was 77.4%, the positive likelihood ratios (PLR) was 3.508, and the negative likelihood ratios (NLR) was 0.267; when the cut-off value of EEG grading was 4.5, the sensitivity was 82.8%, the specificity was 61.3%, the PLR was 2.140, and NLR was 0.281. Conclusions aEEG model was more accurate in prognosticating poor outcomes (3-month) in patients with post-anoxic coma, when compared to EEG grading. Its operation was simple, so aEEG is very suitable in ICU.