Objective To discuss the frequent etiology of spontaneous sub-cortical hemorrhage and its diag- nosis.Methods The clinical materials of 79 cases of spontaneous sub-cortical hemorrhage were analyzed.Results 56% of the hemorrhage was caused by arterial-venous malformation.48% of the hemorrhage was caused by occult AVM.Conclusion AVM is the most frequent etiology of spontaneous sub-cortical hemorrhage.CTA plays a special role in its diagnosis.

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

?AIM: To explore the relation of homocysteine ( Hcy ) and its metabolic related indicators with primary angle-closure glaucoma ( PACG) .?METHODS: In this study, a total of 150 PACG patients and 150 controls were enrolled. The patients with PACG were diagnosed by applanation tonometer, Humphrey perimetry ( HVF ) , optical coherence tomography ( OCT ) , gonioscope. The normal controls were recruited from physical examination center. Blood samples were collected and the plasma was used to determine homocysteine, vitamin B12 and folic acid. DNA was extracted to determine the methylenetetrahydrofolate reductase ( MTHFR ) C677TT genotype and gene polymorphism.?RESULTS:There was a statistically significant difference (t=2. 04,P=0. 04) in mean homocysteine levels between patients with PACG ( 16. 11 ± 1. 66μmol/L ) and controls (15. 74± 1. 52μmol/L). The level of vitamin B12 in PACG group was 248. 09 ± 119. 07pg/mL and the control group was 230. 21 ± 142. 93pg/mL. No statistically significant difference was found on the vitamin B12 level between the two groups ( t = 0. 84, P = 0. 40 ). The folic acid levels showed no statistically significant differences( t=1. 65,P=0.17) between the PACG group(12. 02±7. 81ng/mL) and the control group (13. 15 ± 6. 25ng/mL). The frequency distribution of the MTHFR C677T genotypes was found to be significantly associated (χ2 =6. 2,P<0. 05) with PCAG (CC 84. 7%, CT 4. 7%, TT 10. 7%) as compared to the controls (CC 94. 7%, CT 2. 0%, TT 3. 3%). The frequency distribution of TT genotypes of MTHFR C677T in the PACG group was significantly higher than that in the control group. Moreover, our findings showed significant difference (χ2=14. 2,P<0. 05) between the allele frequency of the C677T single nucleotide polymorphism within the MTHFR gene in POAG patients ( T 87. 0%, C 13. 0%) and the control group(T 95. 7%,C 4. 3%) .?CONCLUSION: These results show that the Hcy level and the TT genotypes of MTHFR C677T were higher in the PACG patients than in controls. Therefore, our data suggests that high Hcy level and the related indicators are associated with high PACG risk.

Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
Amino Acid Sequence ; Carbon-Carbon Double Bond Isomerases ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; Paclitaxel ; biosynthesis ; Phylogeny ; Plant Proteins ; genetics ; Taxus ; enzymology ; genetics

With references of historical materials and modern literature regarding acupoint characteristic, a secondary analysis on the concept, origin, related factors and research methods in present research of acupoint characteristic is performed. The acupoint characteristic should be considered as an acupoint inherent attribute that could explain physiological and pathological manifestations at the same time, including location attribute and function attribute, which is related with time and treatment method. Some re-thinking on acupoint characteristic is proposed as well as advice on further research method and direction, hoping to promote the research development of acupoint characteristic.
Acupuncture Points ; Acupuncture Therapy ; history ; China ; History, Ancient ; Humans ; Medicine in Literature ; Meridians

Autopsy ; Coxsackievirus Infections ; Enterovirus ; isolation & purification ; Enterovirus B, Human ; isolation & purification ; Enterovirus Infections ; Hand, Foot and Mouth Disease ; pathology ; virology ; Humans ; Infant ; Male

OBJECTIVETo study the early change of serum nitric oxide (NO) after acute heat exposure with trauma and the effect of NO on mean arterial pressure (MAP), thus to provide theoretical basis for studying the mechanism of NO effect in acute stress.

METHODSThe rabbit model of acute heat exposure combined with trauma was established. The animals were divided into four groups, including control, trauma, hyperthermia and hyperthermia combined with trauma. The levels of NO were measured at different time points: 0 h, 1 h, 2 h and MAP was monitored throughout the whole experiment.

RESULTSThe concentration of NO declined at first and then increased at 1 h or so after acute heat exposure and trauma. The levels of NO in hyperthermia with trauma group at 1 h, 2 h were (42.75 +/- 8.24), (59.54 +/- 9.05) micro mol/L respectively (P < 0.05), while those in control group were (56.63 +/- 3.79) and (55.22 +/- 7.15) micro mol/L, the difference at 1h between two groups was significant (P < 0.05). Under the circumstance of hyperthermia and trauma, the level of MAP declined to the lowest point at 60 - 70 min and then showed a transient rise, after that, the level declined rapidly.

CONCLUSIONSAt the early stage of acute heat exposure and trauma, the concentration of serum NO declined at first and then increased, and had certain relationship with the change of MAP.

Animals ; Blood Pressure ; Cytokines ; biosynthesis ; Hot Temperature ; Male ; Nitric Oxide ; blood ; Rabbits ; Wounds and Injuries ; blood ; physiopathology

Objective: To explore the expression and clinical significance of serum long chain non-encoded RNAANRIL (LncRNA ANRIL) in chronic heart failure (CHF) patients. Methods: Our research included in 2 groups: CHF group, n=120 patients treated in our hospital and Control group, n=28 healthy subjects at the same period of time. Based on NYHA classification, CHF patients were further divided into 4 subgroups: NYHAⅠ subgroup, n=28, NYHA Ⅱ subgroup, n=34, NYHA Ⅲ subgroup, n=35 and NYHA Ⅳ subgroup, n=23. Expressions of serum ANRIL and cystatin C were examined by real time fluorescence quantitative PCR (QRT-PCR) and compared between 2 groups; the relationship between ANRIL, cystatin C and cardiac function were studied. Results: Compared with Control group, CHF group had increased serum levels of ANRIL and cystatin C, P<0.05. ANRIL expression was gradually increasing by cardiac function decreasing in CHF I to CHF Ⅳ subgroups, P<0.05. Correlation analysis found that serum level of ANRIL was positively related to cystatin C (r=0.873, P<0.001). ANRIL was elevated by increased left ventricular end diastolic diameter and decreased ejection fraction, both P<0.05. Conclusion: Serum ANRIL level has been related to CHF at certain degree which may indirectly reflect the severity of HF in relevant patients.

Objective To evaluate the infectivity and virulence variation caused by mutations in the 5' untranslated region(5'UTR)pyrimidine-rich tract of coxsackievirus B1(CVB1)genome.Methods Five pyrimidines in the 5'UTR pyrimidine-rich tract(nt563-nt573)of CVB1 genome were substituted with purines by site-directed mutagenesis.The mutant,CVB1/m563-573,was purified by plaque assay,and subjected to infectivity and virulence assessments by means of cytopathic effect(CPE),plaque forming,one-step growth curve,and 50% lethal dose(LD50)assays.Results Sequencing data revealed that the sequence of pyrimidine-rich tract in the 5'UTR of CVB1/m563-573 mutant was exactly identical to our design(C565A,U567C,U568A,U570A,and U572G).CPE assay showed that the infectivity of CVB1/m563-573 was weaker than that of its prototype CVB1/wt(A490=0.710±0.074,0.812±0.092)though no significant difference could be observed(t=-2.204,P>0.05).Plaque forming assay showed that the plaque quantities of CVB1/m563-573 were(6.40±1.52)×103,(11.60±2.19)×103 pfu/L and the plaque diameters of CVB1/m563-573 were(2.00±0.35),(2.47±0.41)mm at 46 and 58 hours pestinfection,respectively.The plaque quantities of CVB1/wt were(8.40±2.51)×103,(11.80±1.92)×103 pfu/L and the plaque diameters of CVB1/wt were(1.80±0.27),(2.85±0.44)mm,respectively.There was no significant difference between the plaque quantities and sizes of CVB1/m563-573 and CVB1/wt(t=8.000,0.985,10.000,9.000,all P>0.05).One-step growth curve demonstrated that the numbers(lg)of CVB1/m563-573 progenies at time-points of 3,5,7 h postinfection were 2.10±0.09,4.28±0.03,7.44±0 and that of CVB1/wt progenies were 2.80±0.02,4.77±0.02,8.55±0.01,respectively.The replication of CVB1/m563-573 was significantly slower than that of CVB1/wt at all three time-points(t=-13.151,-24.319,-47.714,all P<0.01).The LD50 of CVB1/m563-573(3.10×109 pfu/L)and CVB1/wt(1.26×107 pfu/L)indicated that the virulence of CVB1/m563-573 was significantly weakened compared to that of CVB1/wt.Conclusions The infectivity and virulence of CVB1 are weakened by substitution of pyrimidines with purines in the pyrimidine-rich tract of CVB1 5'UTR.Site-directed mutagenesis in the pyrimidine-rich tract may be a strategy for developing attenuated CVB vaccine.

OBJECTIVETo study the major metabolites of antitumor lead compound T-OA (oleanolic acyl-3, 5, 6-trimethyl pyrazine-2-methyl ester) in rat urine, in order to preliminarily infer its metabolic mode in rats.

METHODRat urines of the blank group, the raw material group (ligustrazine TMP and oleanolic acid OA Moore equivalent) and the T-OA group were collected and freeze-dried; Solids were extracted by ethyl acetate; And then the extracts were re-dissolved with acetonitrile. HPLC-HRMS coupling technique was adopted to find the potential mass spectrum peak under ESI(+) (see symbol) ESI(-) modes. Metabolite-related information was obtained by comparing the three groups of spectra.

RESULTOne metabolite of OA and two metabolites of TMP were identified in the raw material group; none metabolite of T-OA but one phase II metabolite was detected in the T-OA group.

CONCLUSIONIt is the first time to identify one phase II metabolite of T-OA and one phase II metabolite of OA were identified in rat urine. On that basis, the researchers preliminarily inferred that T-OA does not show the efficacy in the form of raw material. The HPLC-HRMS method established could be used to identify metabolites of related derivative structures. This paper could also provide certain reference for designing pro-drugs based on oleanolic acid.

Animals ; Antineoplastic Agents ; chemistry ; metabolism ; urine ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Male ; Mass Spectrometry ; methods ; Molecular Structure ; Rats ; Rats, Sprague-Dawley