Objective To approach a semi-quantitative diagnostic criteria of neonatal hypoxic-ischemic encephalopathy(HIE) by comprehensive scoring, including history, clinical manifestation and laboratory data,which can be used for early diagnosis,severity grading and the institution of therapy.Methods We analyzed history, clinical manifestation and some parameters of laboratory data,and established criteria for scoring and grading,and compared this grading with the grading determined by clinical data and follow-up,which brought forward a semi-quantitative diagnostic criteria, and applied this criteria to other 41 patients with HIE to test its reliability.Results The grading criteria were established as the following:if total score ≥20.0,the patient was graded as severe one;total score between 10.5-19.5, moderate one; between 5.5-10.0,mild one;≤5.0,non-HIE one. The scoring system was used in other 41 patients with HIE,the specificity and sensitivity were 100% and 96 .9%,97.2% and 100%,100% and 100%, respectively.Conclusions For severe HIE cases who are not permitted to receive compated tomography(CT) examination or no CT can be used, this scoring system can be used to diagnoze HIE and grade the severity. This method is simple and easy to perform.Both specificity and sensitivity are high for diagnosis and severity judgment of HIE.

Aging ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism ; Testis ; drug effects ; metabolism

Background Dominant eye is one of the functional asymmetric organ,and the dfference between dominant eye and undominant eye is a researching hotspot.But the study about accommodation in adult myopia is less.Objective This study was to determine the association between ocular dominances and accommodative factors in the subjects with adult myopia.Methods This study used prospective descriptive research method.Thirty-five subjects aged from 18 to 35 years with the myopia ranged from-2.00 D to-10.00 D and anisometropia less than 1.5 D,BCVA≥ 1.0 were recruited consecutively in this study.Ocular dominance was determined using the hole-inthe-card test and thumb test.Refractive error was measured with objective and subjective optometry,and amplitude of accommodation was measured by push-up test.Fusion cross cylinder(FCC) was used to measure the accommodative lag,and flipper test was applied to determine the accommodative facility.Oral informed consent was obtained from each subject before any relevant examination.Results No significant differences were found in the amplitude of accommodation (D),accommodative facility (cpm) and accommodative lag (D) between the dominant eye and undominant eye (accommodative amplitude:9.69 D±2.30 D vs.9.60 D±2.37 D,P =0.294 ;accommodative facility: 11.08 D±4.20 D vs.10.63 D± 4.60 D,P=0.260;accommodative lag:P=0.141).In the patients with the right eyes as dominance eyes,the accommodative amplitude of both eyes were (9.48±2.29) cpm and (9.33 ± 2.49) cpm,and accommodative facility were (10.50 ± 4.70) cpm and (9.99 ± 4.90) cpm.There were no significant differences between the right and left eyes in the accommodative amplitude,accommodative facility and accommodative lag (P =0.319,0.116,0.590).In the patients with the left eyes as dominant eyes,the accommodative amplitude of both eyes were (9.91±2.35)D and (9.88±2.26) D,and accommodative facility were (10.70±3.77)cpm and (11.25 ±4.27) cpm.No significant differences were seen between the right eyes and left eyes in the accommodative amplitude,accommodative facility and accommodative lag (P =0.749,0.295,0.238).Conclusions The amplitude of accommodation of the dominant eye is not significantly enhanced,and less accommodative lag and better accommodative facility also are found in the demonstrate eye in myopia adults with low anisometropia.

The aim of the present study was to investigate the effect of lipoxin A4 (LXA4)pretreatment on cognitive function of aged rats after global cerebral ischemia reperfusion,and to explore its possible mechanism.Thirty-six aged male Sprague-Dawley rats were randomly divided into three groups (n=12 each):sham-operation group (S group),global cerebral ischemia reperfusion group (I/R group) and LXA4-pretreatment group (L group).The rat model of global cerebral ischemia reperfusion was established by occlusion of the bilateral common carotid artery with hypotension.The cognitive function of rats was determined by a step-down type passive avoidance test and Morris Water Maze test on the third day after reperfusion.Rats were sacrificed after Water Maze test and the pathological changes ofhippocampal CA1 region were observed and the related inflammatory mediators were determined.As compared with S group,the escape latency in I/R group was prolonged from the first day to the fifth day,while that in L group was prolonged from the first day to the third day.The retention time in I/R group and L group in the first quadrant was shortened.The reaction time,frequency of reaction mistake and frequency of escape mistake in I/R group increased,and the latent period shortened.The frequency of escape mistake in L group increased,and the damage in the hippocampal CA1 region of I/R group and L group was obvious.The levels of S-100β,TNF-α,IL-lβ,IL-10 and NF-κB in I/R group and L group increased.As compared with I/R group,the escape latency in L group was shortened from the first day to the fifth day,and the retention time in the first quadrant prolonged.The reaction time,frequency of reaction mistake and frequency of escape mistake in L group decreased,and the latent period prolonged.The damage in the hippocampal CA1 region of L group was alleviated as well.The levels of S-100β,TNF-α,IL-1β and NF-κB in L group decreased,and those of IL-10 increased.It can be concluded that LXA4 pretreatment can improve the cognitive function in aged rats after global cerebral ischemia reperfusion probably by inhibiting the inflammatory reaction.

Objective To observe the effect of T-2 toxin on growth and development of rat epiphyseal plate of left knee and metaphyseal bone of femur and tibia in normal and low nutritional status, to find out possible pathogenic factors of Kashin-Beck disease and provide experimental basis for early intervention. Methods Ninety 3-week-old Wistar rats, weighing 60 - 70 g, were randomly divided into three groups: control group(general feed), T-2 toxin + general feed group, T-2 toxin + low nutrition feed group, thirty rats in each group with equally sex ratio. T-2 toxin (1.0 mg/kg) was administered orally 5 times a week via a gavage needle for 4 weeks. The change of hair, activity and body weight was observed. After 1, 2, 4 weeks, the epiphyseal plate of left knee and metaphyseal bone of femur and tibia (including distal femur and proximal tibia) were collected. Specimens were processed with HE and Masson staining. The morphology of chondrocytes and matrix collagen content in epiphyseal plate was observed. Trabecular bone volume fraction in tibial metaphyseal bone was analyzed by Image-Pro Plus 6.0 software. Results In the control group, rats were in good movement and hair with light, but in T-2 toxin + general feed group and T-2 toxin + low nutrition feed group, rats were found with reduced activities and hair with dark color. Body weights(g) of the control group, the T-2 toxin + general feed group and the T-2 toxin + low nutrition feed group were 81.0 ± 6.2, 79.0 ±5.1, 77.0 ± 7.5, respectively, by the end of first week; 101.8 ± 6.7, 97.0 ± 6.8, 93.0 ± 5.3, respectively, by the end of second week; 151.1 ± 15.7, 126.5 ± 11.9, 106.5 ± 11.5, respectively, by the end of fourth week. There was significant difference in groups by second week and the fourth week (F = 9.72, 41.65, all P ＜ 0.05 ). There was significant difference among multi-groups by the fourth week(all P ＜ 0.01 ). Under light microscope, at the second weeks, coagulative necrosis of chondrocytes was found in hypertrophic zone in the two groups with T-2 toxin; at the fourth weeks, cell necrosis increased. Masson staining showed collagen staining in the two groups with T-2 toxin significantly turned to clear pale coloration, indicating that the collagen matrix was significantly reduced. Image analysis showed there was significant difference in groups at the second and fourth week(F= 9.72, 41.65, all P＜ 0.05)in tibial metaphyseal trabecular bone volume fraction. There was significant difference between T-2 toxin + low nutrition feed group[(0.55 ± 0.12)％, (0.21 ± 0.0)％] and control group[(0.67 ± 0.09)％, (0.51 ± 0.14)％] by the second and fourth week(all P ＜ 0.01 ). Conclusions Under normal nutritional status, T-2 toxin can induce hypertrophic epiphyseal cartilage necrosis, collagen content decreased in epiphyseal plate, metaphyseal trabecular bone formation disorders; in the low nutritional status, T-2 toxin can lead to rat epiphyseal necrosis and significant metaphyseal bone disorder, but whether the performance is related to Kaschin-Beck disease needs to be studied further.

In order to obtain an adequate supply of alpha-galactosidase for research and practical use, the fermentation, purification and identification of the recombinant coffee bean a-galactosidase were carried out. Baffled flasks containing 100mL BMGY were inoculated with the pPIC9K-Gal/GS115 strain and allowed to grow at 30 degrees C, 250- 300r/min until a maximum optical density at 600nm (OD600) between 2.0 to 6.0 was attained. Entire 400 mL seed culture was transferred aseptically to the 5-liter fermenter, which contained 4 liter sterilized basal salts medium and 4% glycerol. The batch culture grew at 30 degrees C, pH 5.0 until the glycerol was completely consumed, and a glycerol feed was initiated to increase the cell biomass prior to induction with methanol. The culture was centrifuged at 8000 x g and the supernatant was collected. Following ultrafiltration, the retentate was balanced in 20 mmol/L sodium formicate buffer, pH 3.8 and loaded onto a cation-exchange column, HiTrap SP. The column was washed with the same buffer and bound proteins were eluted with 1 mol/L NaCl. The fractions containing recombinant a-galactosidase were pooled and concentrated with PEG20 000. Subsequently, the biochemical properties of the enzyme were determined with typical methods. At last, the fresh human blood A and B erythrocytes were incubated with the purified alpha-galactosidase at 26 degrees C for 2 4 hours. Hemagglutinins were assayed by the standard method. After an elapsed fermentation times (EFT) of 18h, the fed-batch phase was initiated to increase the cell biomass. A cellular yield of nearly 200 g/liter wet cells was achieved when induction was initiated. 72h later, the alpha-galactosidase activity against artificial substrate PNPG (PNP-alpha-galactopyranoside) achieved 36 000u per liter culture. The crude fementation supernatant contained few impurities as detected by SDS-PAGE. The supernatant was purified by cation-exchange chromatography, the target alpha-galactosidase was eluted with 40% 1mol/L NaCl and showed a 41kD band on SDS-PAGE. After concentration, the final recovery was about 41%. The Michaelis constant of the recombinant alpha-galactosidase was determined as 0.275 mmol/L, which slightly lower than the nature enzyme and suggested a higher affinity with specific substrate. When human blood type B erythrocytes pretreated with 100u/mL recombinant alpha-galactosidase reacted with bood type B antiserum, no hemagglutination occurred. This suggested that the B antigens had been removed by the enzyme successfully. These results demonstrated that the recombinant alpha-galactosidase could be produced in largescale and made it possible to explore the application of alpha-galactosidase in more fields.
ABO Blood-Group System ; immunology ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Erythrocytes ; drug effects ; Fermentation ; physiology ; Hemagglutination ; drug effects ; Humans ; Pichia ; genetics ; metabolism ; alpha-Galactosidase ; genetics ; metabolism ; pharmacology

Here in this paper introduced is an analysis system for X-ray cephalometry developed through Windows platform,which includes three parts: data input, cephalometry analysis & calculation, and display & printing of diagnostic results. It has been applied in orthodontic department of Stomatological Hospital of the Fourth Military Medical University, and it is quite convenient for doctors' cephalometry diagnosis.
Cephalometry ; instrumentation ; Computers ; Humans ; Medical Records Systems, Computerized ; instrumentation ; Orthodontics ; instrumentation ; methods ; Radiographic Image Enhancement ; instrumentation ; Radiography ; instrumentation ; Software

OBJECTIVETo observe the cell apoptosis after tractive spinal cord injury in rats, determine expression of apoptosis correlative genes, and study the molecular mechanism of cell apoptosis.

METHODSThe T(13)-L(2) spinal cord of rats was injured by traction after the amplitude of P1-N1 wave decreased to 70% in postoperation than in preoperation through cortical somatosensory evoked potential (CSEP) monitor. Then rats were killed in 30 min, 6 h, 1, 4, 7, 14 and 21 d respectively after operation (n = 4). Cell apoptosis was examined by the flow cytometer and terminal deoxynucleotidyl transferase-mediated DUTP-biotin nick end labeling (TUNEL) reaction, the expression of p53, bax and bc1-2 genes was tested with immunohistochemistry.

RESULTSThe flow cytometer test and TUNEL method showed that the apoptosis cell ratio raised in 6 h and reached at peak in 7 d after injury, and then declined till 21 d, they showed significant difference (P < 0.05, 0.01). TUNEL method showed that injured group had a large number of apoptosis glial cells in white matter. Immunohistochemical staining showed that the positive expression of p53, bax and bc1-2 protein raised at 6 h, expression of p53 protein reached at peak in 4 d, bax and bc1-2 protein reached at peak in 7 d after injury. Compared with control group and laminectomy group, the injured group showed significant difference (P < 0.05, 0.01).

CONCLUSIONThere is cell apoptosis phenomenon after tractive spinal cord injury in rats. Morphology indicates that apoptosis includes neurons and glialcytes, which is an important form of cell death and pathological changes in secondary lesion period after tractive spinal cord. There exist high expression of apoptosis correlative gene p53 and bax after spinal cord injury, they may play an important role in reduction of cells to apoptosis.

Animals ; Apoptosis ; genetics ; Disease Models, Animal ; Female ; Gene Expression ; Genes, bcl-2 ; genetics ; Genes, p53 ; genetics ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; genetics ; pathology ; bcl-2-Associated X Protein

Objective To observe the changes in retinal structure after shortpulse laser technique and conventional laser surgery.Methods Oburg fundus photography,spectral domain optical coherence tomography (SD-OCT) and fundus autofluorescence (FAF) examination were performed in 16 patients (22 eyes) receiving shortpulse laser surgery,following 19 patients (25 eyes) undergoing conventional laser surgery to observe the spot situation with the time for postoperative 1 h,1-2 weeks,1-3months,＞3-6 months,＞ 6-12 months.Results After 1 h of short-pulse laser,fundus color displayed light spot of the center was white and gray,which surrounded by gray dizzy;SD-OCT images showed spot ellipsoid area formed a ring out of band,but the inner retina and retinal pigment epithelium (RPE) had no significant change.FAF showed multiple spontaneous fluorescence signals to reduce the circular area;at 1-2weeks after laser surgery,fundus photography showed spot center for white and gray,and the surrounded gray dizzy became larger,and FAF spontaneous fluorescence signal in light spot center was enhanced,while SD-OCT showed that the outer nerve sensory layer was pulled to the photocoagulation spot center.From 1 month to 3 months,some of the epithelial layers of the nerves were restored to normal,and ＞ 3-6 months,the epithelial layer of the nerve was basically restored to normal;and ＞ 6-12 months,the nerve epithelium was back to normal.As for conventional laser surgery,1 h later,fundus photography and FAF performance were similar to the short-pulse laser,and the SD-OCT showed that the whole retinal layer had dropsy in the spot place;after 1-2 weeks,the spot center was gray and white,and the surrounded gray halo turned enlargement,and FAF spontaneous fluorescence signal in light spot center was enhanced,while SD-OCT showed that the outer nerve sensory layer was pulled to the spot center with adhesion;after 1-3 months,fundus photography and FAF performance were similar to those of short-pulse laser,while SD-OCT presented the RPE cells hyperplasia and ring atrophy around the spot,and the RPE atrophy around the spot was gradually enlarged,plus the whole layer of the nerve epithelium was adhesion ＞ 3-6 months after surgery.＞6-12 months later,the results showed the RPE layer atrophy and nerve epithelium layer adhesion.The results of FAF in all follow-up groups were consistent with that of OCT results.Conclusion OCT and FAF are the important methods to observe the postoperative retinal laser structure changes,which can provide objective basis for the confirmation that short-pulse laser has less damage than the conventional laser treatment,and this provides a new research idea to optimize the laser parameters.

This study aimed to assess the relationship of OAS2 rs739901 5'-flanking C/A polymorphisms with the susceptibility to Enterovirus-71 (EV71) infection.We investigated 294 hand-foot-mouth disease (HFMD) Chinese children with EV71 infection (165 mild cases and 129 encephalitis cases).The improved multiplex ligation detection reaction (iMLDR) technique was used to test the genotypes.In EV71-infected patients,the CA genotype distribution (P=0.007),A allele frequency (OR 1.32,95％ CI 1.0-1.7,P=0.034)and CA+AA carriage frequency (P=0.003) of OAS2 rs739901 5'-flanking were obviously elevated as compared with controls,but there were no statistically significant differences between mild cases and encephalitis cases.In EV71-infected patients,the counts of white blood cells (P=0.034) and blood glucose concentrations (P=0.042) were raised in A carriers (CA+AA).Among different genotypes of encephalitis cases,the contents of cerebrospinal fluid (CSF) showed no significant differences.IFN-γ levels in EV71-infected patients were higher than those in controls (mild group vs.control group,P＜0.01;encephalitis group vs.control group,P＜0.001).In encephalitis cases,IFN-γ levels were reduced (P＜0.05) in A carriers compared to CC genotype,however,there were no significant differences between genotypes CA and AA (P=0.226).These findings suggest that OAS2 rs739901 5'-flanking C/A genetic polymorphisms involve the susceptibility to EV71 infection,and A allele might be a risk factor of the susceptibility to EV-71 infection.