Objective To investigate the changes of expression of Fas-associated proteins named Fas-associated death domain protein(FADD)and death-associated protein(Daxx)in the ischemic penumbra following transient focal cerebra ischemia in rats.Methods ①Adult male Sprague-Dawley rats were randomly divided into the sham-operated group and the cerebral ischemia model group.Rats underwent right middle cerebral artery occlusion(MCAO)for 2 h and reperfusion for 1,3,6,12 and 24 h using an intraluminal suture technique.The expression of FADD and Daxx mRNA and protein were measured with methods of immunohistochemistry.Western blot and reverse transcription-polymerase chain reaction(RT- PCR)respectively were used in the ischemic penumbra of rats.②Double-label fluorescence confocal laser scanning microscopy(CLSM)was performed to monitor FADD and Daxx intracellular location before and after ischemia.Results RT-PCR,Immunohistochemistry,Western blot experiments indicated that a very low level of FADD mRNA and protein were detected in the cerebral cortex of sham rats.The expression level both of FADD mRNA and protein increased significantly at 3 h after reperfusion,peaked at 12 h,then declined markedly at 24 h in the ischemic penumbra of model rats.RT-PCR,Immunohistochemistry indicated that a relatively high level of Daxx mRNA was detected in the cerebral cotex of sham rats.The expression level of Daxx mRNA increased significantly at 3 h after reperfusion and persisted to 24 h at a high level,whose protein had a same change of expression level in the ischemic penumbra of model rats. Immunofluorescence double-staining laser scanning by CLSM showed that the immunoreactivity of FADD was located in cytoplasm,and the intracellular translocation of the immunoreactivity of Daxx from nucleus to cytoplasm was monitored by measuring the green fluorescence after ischemia.Conclusion The transient upregulation of FADD and the persistant high level of expression of Daxx may contribute to neuronal apoptosis following cerebral ischemia/reperfusion.

Objective To investigate prognostic factors for bloodstream infection in adult patients. Methods Clinical data of 131 adult patients with positive blood cultures during January 2002 to December 2003 in the Hospital were collected and 91 cases of them were retrospectively analyzed to understand their pathogen species and prognostic factors for it.Results Blood samples from 91 patients were cultured positive,53 cases(58.2%)with gram-negative bacteria mainly including Escherichia coli,Salmonella spp. and Klebsiella pneumoniae,28(30.8%)with gram-positive bacteria,mainly including Staphylococcus aureus and coagnlase-negative Staphylococci,eight(8.8%)with fungi and two(2.2%)with multiple infections.Case fatality ratio in this group of patients with septicemia was 30.8% during their hospitalization,and that in those with Pseudomonas aeruginosa,Staphylococcus aureus,Stenotrophomonas maltophilia and E.coli with extended-spectrum beta-lactamase was over 50%.Case fatality ratio was associated with severity of sepsis(OR=1.15)and inappropriately initial empirical treatment with antibiotics (OR=6.77).Conclusions Pathogen causing bloodstream infection in adults were mainly gram-negative bacteria and severity of infection and inappropriate initial antibiotics treatment could increase their fatality.

Objective To explore the relative factors with the accidents in the cerebral palsy rehabilitation wards, and preventive measures.Methods The accident cases in the rehabilitation wards were recorded respectively.Results There was a relationship between the rehabilitated patient's accident and their parents, the patients themselves, the surroundings and the staff's consciousness of safety.Conclusion Building the safe management organization, strengthening the safety health education, promoting the surroundings in the wards and training for the staff can reduce the accident rate effectively in the rehabilitation wards.

Abstract: Objective　To investigate the diagnostic efficacy of rifampin-resistant real-time fluorescent quantitative nucleic acid amplification detection technology (GeneXpert MTB/RIF) in bronchoalveolar lavage fluid (BALF) combined with peripheral blood tuberculosis infection T cell spot test (T-SPOT) and tuberculosis antibody (TB-Ab) in smear-negative pulmonary tuberculosis. Methods　The clinical data of 114 cases of clinically diagnosed smear-negative pulmonary tuberculosis, 80 cases of non-tuberculous pulmonary diseases and 22 cases of smear-positive pulmonary tuberculosis in our hospital from January 2019 to January 2021 were retrospectively analyzed. The detection results of peripheral blood T-SPOT, TB-Ab and BALF GeneXpert in the three groups were analyzed. The sensitivity, specificity, negative predictive value, positive predictive value, false negative rate, false positive rate and Youden index of the three detection methods were compared. The differences in the positive detection rate of smear-negative pulmonary tuberculosis between the separate detection and the combined detection of the three methods were compared. The receiver operating characteristic curve (ROC) was performed to calculate the area under the curve (AUC). Results　The sensitivity of BALF GeneXpert and peripheral blood T-SPOT and TB-Ab was 66.91%, 80.88% and 90.44%, respectively. The specificity was 98.75%, 73.75% and 41.25%, respectively; the diagnostic coincidence rates were 78.70%, 78.24% and 72.22%, respectively, which were higher than 70.00%. In the smear-negative pulmonary tuberculosis group, the positive detection rates of these three methods in the smear-negative pulmonary tuberculosis group were 63.15%, 79.82% and 90.35%, respectively, and the differences were statistically significant compared with those in the non-tuberculosis pulmonary disease group (all P<0.01). The positive detection rate of the three combined methods in the smear-negative pulmonary tuberculosis group was 96.49 %, which was significantly higher than that of TB-GeneXpert method and T-SPOT, and the differences were statistically significant (χ2=37.283, P<0.01; χ2=13.612, P<0.01); the Youden index of combined detection was significantly higher than that of single detection, and the AUC of combined detection was 0.977, which was significantly higher than that of single detection. Conclusion　BALF GeneXpert combined with peripheral blood T-SPOT and TB-Ab can significantly improve the diagnostic rate of bacterial-negative pulmonary tuberculosis, providing a strong basis for guiding clinical treatment.

Background Elevated intraocular pressure leads to the loss of retinal ganglion cells and vigorous reaction of retinal glial cells.The expression of nestin in retinal glial cells secondary to hypertention and its significance are unclear.ObjectiveThis study aim to investigate the expression of nestin in retinal glial cells (RGCs) in ocular hypertention rats.Methods The ocular hypertention models were established by cauterizing the limbus-draining veins in the right eyes of 42 SD rats,and a conjunctival incision in the left eyes of the rats served as the sham group.The intraocular pressure (IOP) was measured with the Tono-Pen XL tonometer.The number of RGCs in the rats with ocular hypertention was counted.The expression of the nestin protein in RGCs was semi-quantitatively analyzed using Western by immunochemistry.Double immunofluorescence was carried out to evaluate the the confocal laser scaning microscope.Results Significant differences were found in the IOP between the model group and the sham group at various time points (P＜0.05).In 1 week to 3 weeks after operation,the number of RGCs significantly declined in the model group compared with the sham group (P＜0.05).Immunochemistry showed that from 2 hours through 1 week after operation,the expression of nestin was gradually enhanced in the model group in comparison with the sham group.Western blot revealed that the expression of the nestin protein reflected a similar tendency to that of immunofluorescence.The increased introcular pressure as manifested by the induced expression of nestin.Immunoelectron microscopy also confirmed the induced expression of nestin especially at their end-feet suggests a potential neuroprotective mechanism in neuronal degeneration.Nestin may be a useful biomarker for retinal injury study.

Objective To establish a new determination method for the measuring of alkaline phosphatase activity (ALP) with p-acetyl phenyl phosphace (PAP-PNa_2) as substrate.Methods With the help of Vital semiautomatic analyzer,researched a continuous-monitoring procedure and set up experimental parameters.Results When using this assay,the wavelength of PAP's absorption was 325 nm and the Km of ALP was 0.376 mmol/L.The molecular extinction coefficient of PAP at 340 nm was 23 390 L?mol~(-1)? cm~(-1) and the concentration of citrate buffer was 0.438 mol/L.During the process,we found that the optimum pH of enzyme was 10.4,and the concentration of substrate was 5.0 mmol/L.The time of linear reaction was 900 seconds,and the linear range was 0-1 110 U/L.Serum total ALP were 63.1-118.3 U/ L(male) and 52.5-89.0 U/L(female),based on results from 60 heath adults.Conclusions The method is practical in its repetition and convenience,saves time and is not liable to be affected by bilirubin in serum.It is especially suited to the use of automatic analyzers.

Background Normal adult optic disc rim width follows inferior ≥ superior ≥ nasal ≥ temporal (ISNT) rule.Analysis of the optic disc structural parameters with Heidelberg retina tomograph (HRT) can understand whether ISNT principle is suitable to other optic disc structure parameters.Objective This study was to investigate the characteristics of optic nerve head structure parameters in normal Chinese adults.Methods The parents of twins who participated Guangzhou Twin Eye Study were enrolled in Zhongshan Ophthalmic Center from 2006 June to 2011 August.All subjects received the routine eye examinations,including visual acuity,slit lamp microscope,direct ophthalmoscope examination,auto-refraction examination and HRT Ⅲ examination.All data from the right eyes were collected and analyzed.HRT values from different quadrants of optical nerve head were compared with one-way analysis of variance,and the multiple comparison between quadrants was carried out by Bonferroni test.The differences of optical nerve head parameters in different gender were analyzed using independent sample t test and Bland-Altman test.Results In this study,1672 eyes of 1672 subjects were included,with the 759 male and 913 female persons and average age of (41.8±5.3)years.No significant differences were seen in the disc area (DA)between the superior and inferior,as well as the nasal and temporal quadrants among the subjects (P＞0.05).In addition,there were not significantly different in the rim area (RA) between the superior and nasal disc as well as the rim volume (RV) among the inferior,superior and nasal disc (P＞O.05).However,significant differences were found in the cup area (CA),cup/disk area ratio (C/DAR),cup volume (CV),mean cupdepth (MCD),maximum cup depth (MxCD),height variation contour (HVC),cup shape measure (CSM),mean retinal nerve fiber layer thickness (mRNFLT) and retinal nerve fiber layer cross section area (RNFLcsa) among the different quadrants (P＜0.001).The local differences from only DA and RA complied with an ordination of the ISNT rule.The values of DA,CA,C/DAR,CV,MCD,MxCD from male individuals were higher,but those of the RV,HVC,mRNFLT RNFLcsa were lower in comparison with female individuals.Nevertheless,all the differences were ＜ 0.1 with a maximal mean difference in DA and CA (both 0.07 mm2).Conclusions The optical nerve head configurations measured by HRT have different distributing characteristics in four quadrants in normal adults.Most parameters show the higher values in the inferior and superior than those in nasal and temporal of disc,but DA and RA meet ISNT rule.The differences of most parameters between male and female subjects can be ignored.

Background Retinal photocoagulation iconsidered to be an effective therapy fodiabetiretinopathy (DR).Fundufluorescein angiography (FFA) iused to determine if panretinal photocoagulation (PRP) ineeded.SometimeFFiunsuitable fosome Dpatients,and funduphotography igenerally applied.However,whethecolofunduphotography can substitute foFFin evaluating Dgrading and advising foadministering the photocoagulation protocol iunclear.Objective Thistudy wato compare the grading outcomebetween colofunduphotography and FFin assessing DR.Methodprospective case serieself-controlled study wadesigned.Thiclinical study waapproved by the EthiCommittee of Shanghai Tenth People'Hospital,and written informed consenwaobtained from each patienprioto entering the series.Five 50° non-stereoscopiphotographwere taken in 238 eyeof 136 patientwho methe criteriof type 2 diabetes,and then the FFwaperformed.The imagewere analyzed based on the 2003 International DiabetiRetinopathy Disease Severity Scale.The comparison of the grading outcomein assessing Dbetween colofunduphotography and FFwaevaluated by the Kappagreementesusing SPS17.0.ResultFunduphotography showed mild non-proliferative D(NPDR) in 17.2％ of case(41/238),moderate NPDin 20.2％ of case(48/238),severe NPDin 27.7％ of case(66/238) and PDeyein 34.9％ of case(83/238).FFshowed mild NPDin 20.2％ of case(48/238),moderate NPDin 17.6％ of case(42/238),severe NPDin 19.3％ of case(46/238) and PDin 42.9％ of case(102/238).The concordance foDgrading between funduphotography and FFwasubstantial (κ =0.653,P＜0.01).In evaluating whetheiwanecessary to administePRP (to severe NPDand PDpatients),good concurrence waobserved between funduphotography and FF(κ=0.812,P＜0.01).ConclusionThe merge image from five 50° non-stereoscopiphotographpresentagood indicatofothe selection of PRP in Dpatientwith type 2 diabetes.However,the physicianshould be cautiouin the evaluation of retinal non-perfusion areand neovascularization by funduphotography.

Background Heidelberg Retinal Tomograph (HRT) has been widely used in clinical diagnosis of glaucomatous optic neuropathy,importantly,the measurement results of HRT may be affected by myopia.Therefore,studying the effect of different refractive status and axial length on structural parameters of the optic disc is important.Objective This study was to evaluate the influence of refractive error and axial length to optic nerve head parameter measured by HRT.Methods The parents who participate in Guangzhou Twin Study were enrolled in this investigation from July 2006 to August 2011.The regular eye examinations,including visual acuity,slit lamp,fundus,auto-refraction,HRT-Ⅲ and intraocular lens (IOL) Master,were performed on all the subjects.Thc data from right eyes of subjects were collected and analyzed.The linear correlation analysis and multiple linear regression analysis were applied to analyze the correlation between HRT optic papilla configuration parameter and equivalent sphere and ocular axis length.Results A pilot prospective series cases observational study was designed.1748subjects were enrolled in this study.The disc area (DA) (P=0.021),rim area (RA) (P＜0.001),rim volume (RV) (P＜0.001),mean retinal nerve fiber layer thickness (mRNFLt) (P＜0.001) and retinal nerve fiber layer cross section area (RNFLcsa) in the high myopia were greater than those of the emmetropic group,with significant difference between the two groups (P＜0.001).However,the cup area (CA),cup/disk area ratio (C/DAR) and cup volume (CV) were less in the high myopic group than those of the emmetropic group (P=0.004,P＜0.001 and P＜O.001),respectively.In the mild-moderatc myopic group,the RV,mRNFLt and RNFLcsa were greater than those of the emmetropic group (all of the P value less than 0.001),but the numbers of DA,CA,C/DAR,and CV were less than those of the emmetropic group,the differences were significant and their P valuables were 0.038,＜ 0.001,＜0.001 and 0.001 respectively.The differences of structure parameters between the hyperopia group and the emmetropic group were not statistically significant (P＞0.05).The DA,CA,RA,C/DAR,CV,RV,mean cup depth (MCD),maximum cup depth (MxCD),cup shape measure (CSM),mRNFLt and RNTLcsa were (2.04±0.43) mm2,(0.49±0.34) mm2,(1.55 ± 0.34) mm2,0.23-0.13,(0.11 ± 0.12) mm3,(0.45 ± 0.19) mm3,(0.21 ± 0.09) mm,(0.60±0.21) mm,-0.21 ± 0.07,(0.29 ± 0.07) mm and (1.46 ± 0.36)mm2,respectively,in the 1748 eyes of 1748 subjects.No significant correlation was found between the DA value with SE (r=-0.032,P =0.186).However,weaker positive correlations were seen between the CA,C/DAR,CV,MCD,MxCD with SE (r=0.139,0.163,0.130,0.004,0.054,P＜ 0.05) and negative correlation between RA,RV,mRNFLt and RNFLcsa with SE (r =-0.178,-0.331,0.241,-0.239,P＜0.001).With the increase of the ocular axial length,the DA,RA,RV,mRNFLt and RNFLcsa values were enlarged,and the CV value was smaller.With the increase of CA,the RA,C/DAR,CV,RV,MCD,MxCD,CSM and RNFLcsa values elevated,but mRNFLt value was decreased.Conclusions Refractive error is not associated with DA,but the measuring results are different among various refractive groups.Refractive error,ocular axial length and DA affect the measuring result of HRT.

Objective To explore the effects of mild and moderate acute hypobaric hypoxia on manual performance.Methods Using hypobaric chamber to simulate hypoxia conditions and devising 4 kinds of objective ergonomic testing items(Insert sticks into holes-board,ISIHB;nut-bolt assembly task,NBAT;shape discrimination,SD;and Grip strength,GS including fatigue and tolerance)and one subjective research item(questionnaire subjective sense)to examine manual work efficiency varieties of 9 subjects exposed to a hypobaric chamber with 5 simulated altitudes(3 500,4 000,4 500,5 000 and 5 500 m),for(25?5)min.Results Compared to control group(50 m,the altitude of Beijing):Accomplish time(AT)performance of ISIHB and NBAT significantly decreased(P