Background Human limbal allograft transplantation or limbal autograft transplantation are the primary approaches to the severe corneal-blindness,but their application in clinic were limited because of the defects of donor material.With the development of tissue engineering technology,transplantation of in vitro cultured limbal epithelial stem cells is being an advanced management.Objective The aim of this work was to expand human limbal epithelial stem cells ex vivo under the guidance of confocal microscope and to lay the foundation for fabricating ex vivo cultured cell sheets.Methods Ten eyes of ten patients were examined with the Heidelberg Retina Tomography Ⅲ Rostock Cornea Module(HRT3-RCM)to elucidate the structure of the human corneoscleral limbus and to correlate limbal epithelial dimensions.According to the analysis of the images of limbal epithelia,the limbal tissues provided by Eye Bank of Henan Eye Institute were cut into suitable explants.Then,this study was conducted to expand limbal epithelial stem cells ex vivo on denuded amniotic membrane.The phenotypes of primary cultured cells were evaluated by morphology and immunofluorescent staining with antibodies for limbal epithelial stem cell markers (p63,cytokeratinl9)and differentiation markers(keratin 3,involucrin).This experimental procedure was approved by the Ethic Committee of Henan Provincial People's Hospital.The written informed consent was obtained from subjects before initiation of any examination.Results The palisade morphology of human limbus was imaged clearly on the laser scanning in vivo confocal microscopy and many hyperreflective cells were observed in palisade basal cells.The cell-island phenomenon was seen in the basement membrane under the laser scanning in vivo confocal microscopy.The oblique sections of limbus showed many papilla-like epithelial columns below the superficial limbal epithelia.Throughout the experiment duration,the epithelial cells grew well with the migration rates from limbal tissue (68.62± 16.94)％ and the migration time(5.83 ±2.04)days,which depended on the tissue freshness.Compared with the second and forth batch of tissue,the migration rates of the third and sixth batch of tissues were significantly higher(P＜0.05),and the migration time was evidently longer in the forth and sixth batch of tissue compared with the first,second,third and fifth batch(P＜0.05).The positively expressing rates in the cultured corneal stem cells were 4.05％ and 36.52％ for p63,26.07％ and 40.55％ for CK19,57.88％ and 40.81％ for K3,64.66％ and 59.19％ for involucrin.Conclusion Human limbal epithelial stem cells can be successfully and purposefully obtained from the limbal tissue based on the guidance confocal miscroscope.The cultured corneal stem cells can grow well on the denuded amniotic membrane

The extracting technology of salidroside, tyrosol, crenulatin and gallic acid from Rhodiolae Crenulatae Radix et Rhizoma was optimized. With extraction rate of salidroside, tyrosol, crenulatin and gallic acid as indexes, orthogonal test was used to evaluate effect of 4 factors on extracting technology, including concentration of solvent, the dosage of solvent, duration of extraction, and frequency of extraction. The results showed that, the best extracting technology was to extract in 70% alcohol with 8 times the weight of herbal medicine for 2 times, with 3 hours once. High extraction rate of salidroside, tyrosol, crenulatin and gallic acid were obtained with the present technology. The extracting technology was stable and feasible with high extraction rate of four compounds from Rhodiolae Crenulatae Radix et Rhizoma, it was suitable for industrial production.
Chemical Fractionation ; methods ; Chemistry, Pharmaceutical ; methods ; Coumarins ; isolation & purification ; Drugs, Chinese Herbal ; isolation & purification ; Gallic Acid ; isolation & purification ; Glucosides ; isolation & purification ; Phenols ; isolation & purification ; Phenylethyl Alcohol ; analogs & derivatives ; isolation & purification ; Rhizome ; chemistry ; Rhodiola ; chemistry

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.

The eight heavy metals and two essential constitutes of safflowers planted in linzhi which lies in Southern Tibet were analyzed by ICP-MS and by HPLC respectively. Heavy metals of safflower in the region were at the lower level and the essential constitutes were at the higher level. The better quality of safflower here was assisted by the excellent climate in tibet.
Carthamus tinctorius ; chemistry ; Chromatography, High Pressure Liquid ; Drug Contamination ; Drugs, Chinese Herbal ; analysis ; Flowers ; chemistry ; Metals, Heavy ; analysis ; Tibet

Acute Disease ; Animals ; Behavior, Animal ; drug effects ; Female ; Male ; Mice ; Mice, Inbred ICR ; Nitriles ; poisoning ; Rabbits

Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; complications ; genetics ; pathology ; Male ; Thymus Gland ; abnormalities

Objective To understand the information of food hypersensitivity of children with Henoch-Schonlein purpura (HSP) by food intolerance test.Methods Seventy-four children (40 boys and 34 girls;aged from 3 to 14) with HSP who were hospitalized in our hospital from Dec.2006 to May 2007 were chosen.Two mils venous blood was collected from each object,separated serum.Enzymelinked immunosorbent assay method was applied to detect the serum concentration of 14 kinds food allergen IgG in 74 children with HSP.The concentration of food allergen IgG was divided into 4 levels: 0(IgG0.20 U/L,servious allergy).Results The positive rate was 77.0% in 74 cases with HSP.One type of food allergen specific IgG increased in 24 cases(42.1%),2 types of antibodies increased in 16 cases(28.1%),3 types of antibodies increased in 12 cases(21.1%).Four or more types of antibodies increased in 5 cases(8.7%).The positive rates of 14 food allergen IgG were as follow:egg 93.0%,milk 26.3%,soybean 15.8%,tomato 14.0%,wheat 12.3%,ling 12.3%,shrimp 8.8%,crab 8.8%,beef 3.5%,corn 1.8%,rice 1.8%.Conclusion Food intolerance test is a useful method for children with HSP to find food allergen and guideline for diet of these children.

AlM:To investigate the prevalence of pterygium of the household population aged 40 and above in Hengli Town of Dongguan.
METHODS: Using the method of cluster random sampling, select 3 628 people aged 40 and above in four villages and one community for visual examination, intraocular pressure check, slit lamp examination and questionnaire.
RESULTS: The actual number of subjects was 3 393 people, and examination rate was 93. 52%. We detected 843 patients with pterygium. The prevalence of pterygium was 24. 85%.
CONCLUSlON:There is high prevalence of pterygium in Dongguan area. The prevalence of pterygium is related with age and working environment, but has no relation with gender.

Objective To investigate the spatial distribution and clustering areas of Budd-Chiari syndrome in Heze City,Shandong Province,and to provide epidemiological information for further exploring the etiology and related risk factors of the disease.Methods Detailed residential addresses of 342 cases of patients (residents of Heze City) with diaphragm type Budd-Chiari syndrome diagnosed between 1995 and 2004 in Heze Municipal Hospital,Heze Shan County Central Hospital,Affiliated Hospital of Xuzhou Medical College,Shandong Provincial Hospital and Beijing Xuanwu Hospital were collected.Geographic information system (GIS) was used as a platform for data management and display.The nearest neighbor index,Ripley's K(d) function,Ripley's L(d) function and the nearest neighbor clustering method were applied to detect the spatial characters of Budd-Chiari syndrome in Heze City,Shandong Province.Crimestat 3.0 was used for spatial analysis.Results The nearest neighbor distance analysis showed that the nearest neighbor index was 0.6767 (Z =-11.4387,P ＜ 0.01).That was an aggregation at the first-order spatial scale.Within the study area,the first clustering radius of Budd-Chiari syndrome was 6.66 km,and the first clustering strength was 5.40; the average radius of the strongest clustering area was 126.61 km,and the clustering strength was 12.52,while the biggest clustering radius was larger than 222 km.After corrected by population,the gathering strength was slightly higher than that before the correction.Ten first-order hot spots were formed,and 95％ confidence interval aggregation number was 7,which meant the results were statistically significant(P ＜ 0.05),main clustering areas are in Mudan District,Shan County and Juancheng.One secondorder hot spot was gathered based on the first-order hot spot.Conclusions Spatial distribution of Budd-Chiari syndrome in Heze City,Shandong Province has showed spatial aggregation and heterogeneity.This study has a great epidemiological significance for further exploring the cause of Budd-Chiari syndrome.

Objective To undemtand the rhythm of urinary iodine level of children aged 8-10 in Chongqing city.Methods In April 2008,using the stratified random sampling method,we sampled 60 children aged 8-10 in a lodging primary school in Chongqing(20 per age group,half male and half female),the urine samples were collected in the morning and at 10:00,12:30,16:00,iodine in urine was detected by method of Ce and arsenic catalytic speetrophotometry(WS/T 107-2006).The difference of the urinary iodine level was compared by age,sex and time of day.Results The median urinary iodine of 60 children was 265.07μg/L on the overall.Irrespective of the stratification factors,excluding morning urinary iodine(366.75μg/L)and urinary iodine at 10:00(338.30 μg/L),the urinary iodine between 12:30(235.15μg/L)and 16:00(251.50μg/L)was not significant(all P>0.05),statistically significant differences(all P<0.05)were found between any two.The urinary iodine of 8-year-old group at different times of the day was significantly different(all P<0.05),except between morning urinary iodine (298.90 μg/L)and at 10:00,16:00(279.00,286.59 μg/L),between urinary iodine at 10:00 and 16:00(all P>0.05).The 9-year-old group's urinary iodine were not significantly different between morning urine(366.15μg/L)and 10:00(368.10 μg/L),and between 12:30(244.00 μg/L)and 16:00(186.30 μg/L,all P>0.05),significant differences were faund at other times of the day(all P<0.05).The 10-year-old group of urinary iodine changed very little before 12:30 (382.85,449.60,337.00 μg/L, all P > 0.05 ), followed by rapid decline to 16: 00 (269.35 μg/L), and compared with the morning urine and 10:00, there was significant difference(all P < 0.05).Regardless boys or girls, the urinary iodine at different times qf the day was significantly different (all P < 0.05),except between morning urinary iodine(337.32,309.28 μg/L) and at 10:00(316.15,288.27 μg/L), between urinary iodine at 12:30(251.18,211.45 μg/L) and 16:00(235.02,211.45 μg/L, all P > 0.05). Conclusions The change of urinary iodine level in children aged 8 - 10 was not obvious before noon, changes can be seen in the afternoon.Urinary iodine level before 10:00 is indicative.