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Aim: To evaluate the potential of two trabecular meshwork (TM)-specific promoters, Chitinase 3-like 1 (Ch3L1) and matrix gla protein (MGP), for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2 (scAAV2) vector technologies., Methods: An scAAV2 vector with C3 transferase (C3) as the reporter gene (scAAV2-C3) was selected. The scAAV2-C3 vectors were driven by Ch3L1 (scAAV2-Ch3L1-C3), MGP (scAAV2-MGP-C3), enhanced MGP (scAAV2-eMGP-C3) and cytomegalovirus (scAAV2-CMV-C3), respectively. The cultured primary human TM cells were treated with each vector at different multiplicities of infections. Changes in cell morphology were observed by phase contrast microscopy. Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining, real-time quantitative polymerase chain reaction and Western blot. Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively. In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope. Intraocular pressure (IOP) was monitored using a rebound tonometer. Ocular responses were evaluated by slit-lamp microscopy. Ocular histopathology analysis was examined by hematoxylin and eosin staining., Results: In TM cell culture studies, the vector-mediated C3 expression induced morphologic changes, disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rho-associated protein kinase signaling pathway. At the same dose, these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3, but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3. At low-injected dose, the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGP-C3-injected and scAAV2-eMGP-C3-injected eyes. At high-injected dose, significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes. Similar to scAAV2-CMV-C3, scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium. In anterior segment tissues of scAAV2-eMGP-C3-injected eyes, no obvious morphological changes were found except for the TM. Inflammation was absent., Conclusion: In scAAV2-transduced TM cells, the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus, but obviously higher than that of MGP. In the anterior chamber of rat eye, the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter, but not by Ch3L1 promoter. These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector., (International Journal of Ophthalmology Press.)

A new species Coomaniella sunfengyii Liao, Su, Qi & Song, sp. nov. from Fujian Province, China, is described and placed in the Coomaniella macropus species-group. The description, illustrations, host plant information and diagnostic characters of the new species are provided.

Aim: To investigate the causal gene mutation and clinical characteristics for two Chinese families with autosomal dominant congenital coralliform cataract., Methods: Two Chinese pedigrees with congenital cataract were investigated. Routine ophthalmic examinations were performed on all patients and non-affected family members. Peripheral blood samples were collected, and the genomic DNAs were extracted. The coding regions of proband's DNAs were analyzed with cataract gene panel. The identified mutation was amplified by polymerase chain reaction, and automated sequencing was performed in other members of two families to verify whether the mutated gene was co-segregated with the disease., Results: Congenital coralliform cataract was inherited in an autosomal dominant mode in both pedigrees. For each family, more than half of the family members were affected. All patients presented with severe visual impairment after birth as a result of bilateral symmetric coralliform lens opacification. An exact the same defect in the same gene, a heterozygous mutation of c.70C>A (p. P24T) in exon 2 of γD-crystallin gene, was detected in both probands from each family. Sanger sequencing analysis demonstrated that the mutated CRYGD was co-segregated in these two families., Conclusion: A c.70C>A (p. P24T) variant in CRYGD gene was reconfirmed to be the causal gene in two Chinese pedigrees. It is known that mutated CRYGD caused most of the congenital coralliform cataracts, suggesting that the CRYGD gene is associated with coralliform congenital cataract., (International Journal of Ophthalmology Press.)

Background: An increase in alcohol consumption and related harmful effects has been reported among the elderly population in Asia. Of note, it is important to monitor patterns of alcohol use, and to establish a valid and reliable evaluation system when screening for risky consumption in this age group., Objective: The aim of the current study was to evaluate the possible alcoholic liver disease (ALD) risk factors of a local population in elderly Chinese adults., Methods: A questionnaire was sent to 3393 Chinese adults over 40 years old in Hefei. Alcohol consumption was determined based on the AUDIT questionnaire. ALD was defined by ALD diagnostic standards. Adjusted odds ratios and 95% confidence intervals (95% CI) derived from multiple logistic regression models were used to assess the relationship between ALD and sociodemographic variables., Results: Among 2545 total interviewees, 448 (17.6%) reported a history of alcohol consumption in the previous 12 months. Of these drinkers, 46.7% were male and 53.3% were female. The overall Cronbach's alpha coefficient for AUDIT was 0.648. The rate of ALD was 6.83%. Alcohol abuse was significantly associated with ALD. In the logistical model, alcohol abuse was independently associated with ALD (OR = 6.17, 95% CI: 3.69-15.24; p < 0.01)., Conclusions: Alcohol use, sex, age, and facial flushing were risk factors for ALD. These results provide important evidence for the prevention and therapy of ALD., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Transmembrane protein 88 (Tmem88) is a crucial inhibitor for Wnt/β-catenin pathway in the development of myocardial cells. Due to the important role of β-catenin in the activation and proliferation of hepatic stellate cells (HSCs), it is necessary to investigate the function of Tmem88 in HSCs. In this study, we found that Tmem88 expression was decreased in the human liver fibrotic tissues, primary HSCs from fibrotic mice and activated HSC-T6 cells. Functionally, Tmem88 could inhibit HSCs activation and proliferation by blocking Wnt/β-catenin pathway, and promoted the apoptosis of activated HSCs by initiating Bcl-2/Bax/Caspase3 pathway. Moreover, the level of DNA metyltransferase 3a (Dnmt3a) was upregulated in activated HSCs, and siRNA-mediated Dnmt3a silencing led to Tmem88 restoration. These results indicated that Tmem88 played an important role in HSCs activation, proliferation and apoptosis, and Tmem88 expression might be modulated by Dnmt3a., (Copyright © 2016. Published by Elsevier Ltd.)

Objective: To explore the dynamic changes of D-dimers during pregnancy and early puerperium(within 3 days postpartum). Methods: A retrospective study was performed among 8 367 healthy women who had term singleton delivery in Women's Hospital, School of Medicine, Zhejiang University from January 2007 to December 2014. D-dimers concentrations during pregnancy and early puerprium of all the cases were collected. Data of 21 065 D-dimers tests were assigned to 5 groups according to the time of sampling, including early pregnancy(≤12 gestation weeks), middle pregnancy(12-28 gestation weeks), late pregnancy(>28 gestation weeks), 1 postpartum(within 48 hours postpartum)and 2 postpartum(48- 72 hours postpartum). The D-dimers concentrations in different groups were compared. The effect of delivery mode on D-dimers of early pureperium was analyzed. The correlation between D-dimers and the thromboembolic disease was also explored. In this study, Student's t -test and Wilcoxon rank sum test were used for statistical analysis. D-dimers concentration ≤0.5 mg/L was used as the normal range. Results: (1)D-dimers concentrations during pregnancy were higher than the non-pregnant women( P <0.01), but there was no statistical difference between early pregnancy and late pregnancy( P =0.820). D-dimers concentration in the 1 postpartum group was higher than that of early pregnancy group or late pregnancy group( P <0.01). But in the 2 postpartum group, it was lower than early pregnancy, late pregnancy and 1 postpartum groups.(2)D-dimers in cesarean section cases was significantly higher than in vaginal delivery cases in each period of pregnancy and early pueprium.(3)The 95% CI of D-dimers in early pregnancy, late pregnancy, 48 hours after vaginal delivery, 48- 72 hours after vaginal delivery, ≤48 hours after cesarean section, 48- 72 hours after cesarean section were 0.58-8.28, 0.47-11.52, 1.04-9.59, 0.87-5.22, 1.07-11.58 and 1.00-6.23 mg/L, respectively.(4)In 6 cases with thromboembolic disease, D-dimers was 6.89- 19.89 mg/L, with the mean value of 13.66 mg/L. It was significantly higher than normal range. In 3 cases, all after cesarean section, with lower extremity vein thrombosis within 48 hours postpartum, the D-dimers concentrations, 9.77, 8.65 and 6.89 mg/L respectively, were in the 95% CI of the study population after cesarean section. Conclusions: D-dimers concentration of 0.5 mg/L is not suitable for venous thromboembolism screening during pregnancy. D-dimers concentration in pregnancy and early puerprium is higher than non-pregnancy. It increases in the very early period postpartum and decreases with time. D-dimers should not be a routine screening test to exclude thromboembolic disease in pregnant women without high risk factors and clinical manifestation of thromboembolic disease.

Liver fibrosis is a reversible wound-healing response to chronic hepatic injuries. Activation of hepatic stellate cells (HSCs) plays a pivotal role in the development of hepatic fibrosis. The currently accepted mechanism for the resolution of liver fibrosis is the apoptosis and inactivation of activated HSCs. Protein tyrosine phosphatase 1B (PTP1B), a prototype of non-receptor protein tyrosine phosphatase, is proved to be a vital modulator in cardiac fibrogenesis. However, the precise role of PTP1B on liver fibrosis and HSC activation is still unclear. Our study showed that the expression of PTP1B was elevated in fibrotic liver but reduced after spontaneous recovery. Moreover, stimulation of HSC-T6 cells with transforming growth factor-β1 (TGF-β1) resulted in a dose/time-dependent increase of PTP1B mRNA and protein. Co-incubation of HSC-T6 cells with PTP1B-siRNA inhibited the cell proliferation and activation induced by TGF-β1. Additionally, both mRNA and protein of PTP1B were dramatically decreased in inactivated HSCs after treated with adipogenic differentiation mixture (MDI). Over-expression of PTP1B hindered the inactivation of HSC-T6 cells induced by MDI. These observations revealed a regulatory role of PTP1B in liver fibrosis and implied PTP1B as a potential therapeutic target., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Phosphorylation of tyrosine residues within proteins, which is controlled by the reciprocal action of protein tyrosine kinases and protein tyrosine phosphatases, plays a key role in regulating almost all physiological responses. Therefore, it comes as no surprise that once the balance of tyrosine phosphorylation is disturbed, drastic effects can occur. Protein tyrosine phosphatase 1B (PTP1B), a classical non-transmembrane tyrosine phosphatase, is a pivotal regulator and promising drug target in type 2 diabetes and obesity. Recently it has received renewed attention in liver diseases and represents an intriguing opportunity as a drug target by modulating hepatocyte death and survival, hepatic lipogenesis and so on. Here, the multiple roles of PTP1B in liver diseases will be presented, with respect to liver regeneration, drug-induced liver disease, non-alcoholic fatty liver disease (NAFLD) and hepatocellular carcinoma., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

The purpose of this study was to identify the clinical features and mutations in the fibrillin-1 gene (FBN1) in a large Chinese family with autosomal dominant Marfan syndrome (MFS). Seventeen members from a Chinese family of 4 generations were included in the study. All members underwent complete ophthalmic examination. Molecular genetic analysis was performed on all subjects. All exons of FBN1 were amplified by polymerase chain reaction, sequenced, and the sequences were compared with a reference database. Variations were evaluated in family members as well as 100 normal controls. Changes in structure and function of the protein induced by amino acid variation were predicted by bioinformatic analysis. Ectopia lentis, dolichostenomelia, arachnodactyly, and tall stature were present in all patients diagnosed with MFS. The novel heterozygous missense mutation c.2243 T>G (p.C781W) in exon 19 of FBN1 was identified in all 5 patients, but not in other family members or 100 normal controls. This mutation caused an amino acid substitution of cysteine to tryptophan at position 781 (p.C781W) of the FBN1 protein. This mutation occurred in a highly conserved region and may cause structural and functional changes in the protein according to our bioinformatic analysis. Our results suggest that the novel mutation C781W of FBN1 is responsible for the pathogenesis of MFS in this pedigree.

The purpose of this study is to evaluate the neuroprotective effects of C3 exoenzyme (C3) on N-methyl-d-aspartate (NMDA)-induced retinopathy in rats. C3 was expressed in Escherichia. coli and purified by affinity chromatography. Immunofluorescence was performed in NIH 3T3 cells treated with C3 to verify the cellular uptake of the protein. NMDA was injected intravitreally into rat eyes with or without C3. At various time points after injection, eyes were enucleated. Hematoxylin/eosin staining was performed on retina cross-sections for morphological analysis. Survival and apoptosis of cells in the ganglion cell layer (GCL) were assessed by cresyl violet staining and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) on retina flat-mounts. RhoA levels in retina cells were evaluated by Western blot to detect C3 uptake in vivo. The cellular uptake of C3 was verified by immunofluorescence. Damage including a decrease in inner plexiform layer (IPL) thickness and reduction of cell density in the GCL, corresponding to apoptosis of neurons, was induced by intravitreal injection of NMDA. Protection against this damage was observed following co-injection of C3 and NMDA. RhoA ADP-ribosylation induced by C3 was confirmed by Western blot. Our results suggest that C3 exerts neuroprotective effects against excitotoxic damage induced by NMDA., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

This study aimed to characterize the clinical features of a Chinese pedigree with neurofibromatosis type 1 (NF1) and to identify mutations in the NF1 gene. In this three-generation family containing 8 members, 5 had been diagnosed with NF1 and the others were asymptomatic. All members of the family underwent complete medical examinations. Molecular genetic analyses were performed on all subjects included in the study. All exons of NF1 were amplified by polymerase chain reaction, sequenced, and compared with a reference database. Possible changes in function of the protein induced by amino acid variants were predicted by bioinformatic analysis. In this family, the 5 patients presented different clinical phenotypes, but all manifested typical café-au-lait macules. One novel frame-shift mutation, c.702&#95;703delGT, in exon 7 of NF1 was identified in all affected family members, but not in the unaffected family members or in 102 normal controls. This mutation generates a premature stop codon at amino acid position 720. Additionally, a synonymous mutation c.702 G>A was found in 3 family members, including 2 affected and 1 normal individuals. In conclusion, our study suggests that a novel c.702&#95;703delGT frame-shift mutation in NF1 is likely to be responsible for the pathogenesis of NF1 in this family. To the best of our knowledge, it is the first time that a c.702&#95;703delGT mutation has been identified in a family with neurofibromatosis type 1.

Aim: To detect the mutations in two candidate genes, myocilin (MYOC) and cytochrome P450 1B1 (CYP1B1), in a Chinese family with primary open angle glaucoma (POAG)., Methods: The family was composed of three members, the parents and a daughter. All members of the family underwent complete ophthalmologic examinations. Exons of MYOC and CYP1B1 genes were screened for sequence alterations by polymerase chain reaction (PCR) and direct DNA sequencing., Results: The mother was the proband, she was diagnosed as POAG in both eyes. Her daughter was diagnosed as juvenile-onset POAG. The father was asymptomatic. One MYOC heterozygous mutation c.1150 G>A (D384N) in exon 3 was identified in the mother, another MYOC heterozygous variation c.1058 C>T (T353I) in exon 3 was identified in the father, and the daughter inherited both of the variations. Meanwhile, three single nucleotide polymorphisms (SNPs) in CYP1B1 gene were found in the family., Conclusion: The D384N mutation of MYOC has been reported as one of disease-causing mutations in POAG, whereas T353I variation of MYOC was thought as a high risk factor for POAG. The two variations of MYOC were first reported in one juvenile-onset POAG patient who presented with more severe clinical manifestations, suggesting that T353I polymorphism of MYOC may be associated with the severity of POAG.

Glaucoma, as characterized by accelerated retinal ganglion cell (RGC) death and cupping of optic nerve head (ONH), is one of the leading causes of blindness worldwide. Elevated intraocular pressure (IOP) is generally considered as a major risk factor in the pathogenesis of glaucoma. Previous studies showed that glaucoma caused decrease in collagen and elastin density in several ocular tissues, such as lamina cribrosa, peripapillary sclera and cornea, and resulted in reduced elasticity and compliance of these tissues. It is known that estrogen has protective effects against glaucoma, yet the underlying mechanism still remains obscure. Prior researches have provided evidences showing that the estrogen receptors (ERs) express in a variety of the ocular tissues. Estrogen activates the synthesis of collagen fiber and improves the compliance of these tissues. This leads to a reasonable postulation that increased estrogen may result in a higher content of the collagen fibers and enhanced flexibility of the whole eye, which would therefore decrease IOP. Particularly, the increase in the amounts of collagen fibers at lamina cribrosa improves its compliance, which in turn relieves its compression on RGC axons. Therefore, even at the same IOP level, the softening of cribriform foramina yields a more flexible environment for the RGCs to survive. We therefore hypothesize that estrogen at proper dosage can be considered as a potential therapy for glaucoma since it is able to prevent the eye from glaucomatous damage and lower IOP, especially for those menopausal women with glaucoma., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: The purpose of this study was to elucidate the molecular basis of ocular albinism type I in a Chinese pedigree., Methodology/principal Findings: Complete ophthalmologic examinations were performed on 4 patients, 7 carriers and 17 unaffected individuals in this five-generation family. All coding exons of four-point-one (4.1), ezrin, radixin, moesin (FERM) domain-containing 7 (FRMD7) and G protein-coupled receptor 143 (GPR143) genes were amplified by polymerase chain reaction (PCR), sequenced and compared with a reference database. Ocular albinism and nystagmus were found in all patients of this family. Macular hypoplasia was present in the patients including the proband. A novel nonsense hemizygous mutation c.807T>A in the GPR143 gene was identified in four patients and the heterozygous mutation was found in seven asymptomatic individuals. This mutation is a substitution of tyrosine for adenine which leads to a premature stop codon at position 269 (p.Y269X) of GPR143., Conclusions/significance: This is the first report that p.Y269X mutation of GPR143 gene is responsible for the pathogenesis of familial ocular albinism. These results expand the mutation spectrum of GPR143, and demonstrate the clinical characteristics of ocular albinism type I in Chinese population.

Purpose: To characterize the clinical features of a Chinese Uygur pedigree with primary open-angle glaucoma (POAG) and to identify mutations in two candidate genes, trabecular meshwork inducible glucocorticoid response (MYOC/TIGR) and human dioxin-inducible cytochrome P450 (CYP1B1)., Methods: Twenty one members from a Chinese Uygur family of four generations were included in the study. All participants underwent complete ophthalmologic examinations. Five were diagnosed as POAG, four as glaucoma suspects, and the rest were asymptomatic. Molecular genetic analysis was performed on all subjects included in the study. All exons of CYP1B1 and MYOC were amplified by polymerase chain reaction (PCR), sequenced and compared with a reference database. The variations detected were evaluated in available family members as well as 102 normal controls. Possible changes in structure and function of the protein induced by amino acid variance were predicted by bioinformatics analysis., Results: Elevated intraocular pressure and late-stage glaucomatous cupping of the optic disc were found in five patients of this family. A novel heterozygous missense mutation c.1151 A>G in exon 3 of MYOC was found in all five patients diagnosed as POAG and four glaucoma suspects, but not in the rest of the family members and 102 normal controls. This mutation caused an amino acid substitution of aspartic acid to glycine at position 384 (p. D384G) of the MYOC protein. This substitution may cause structural and functional changes of the protein based on bioinformatics analysis. No mutations were found in CYP1B1., Conclusions: Our study suggests that the novel mutation D384G of MYOC is likely responsible for the pathogenesis of POAG in this pedigree.

Aim: To train Tibetan monkey (Macaca thibetana) for intraocular pressure (IOP) measurement in conscious state and obtain normal IOP in conscious Tibetan Macaque., Methods: The training was based on award-conditioned behavior. Food stimulation and human-animal interaction were used in this training., Results: Trained Tibetan monkeys calmly accepted IOP measurement by the TonoVet® rebound tonometer without sedation or anesthesia and their IOP values were similar to other primates., Conclusion: Human-cultivated Thibetan monkeys are tamable, and can be used for biomedical research such as ophthalmic research without anesthesia.

Retinitis pigmentosa (RP) is a heterogeneous group of progressive retinal degenerations characterized by pigmentation and atrophy in the mid-periphery of the retina. Twenty two subjects from a four-generation Chinese family with RP and thin cornea, congenital cataract and high myopia is reported in this study. All family members underwent complete ophthalmologic examinations. Patients of the family presented with bone spicule-shaped pigment deposits in retina, retinal vascular attenuation, retinal and choroidal dystrophy, as well as punctate opacity of the lens, reduced cornea thickness and high myopia. Peripheral venous blood was obtained from all patients and their family members for genetic analysis. After mutation analysis in a few known RP candidate genes, exome sequencing was used to analyze the exomes of 3 patients III2, III4, III6 and the unaffected mother II2. A total of 34,693 variations shared by 3 patients were subjected to several filtering steps against existing variation databases. Identified variations were verified in the rest family members by PCR and Sanger sequencing. Compound heterozygous c.802-8&#95;810del17insGC and c.1091-2A>G mutations of the CYP4V2 gene, known as genetic defects for Bietti crystalline corneoretinal dystrophy, were identified as causative mutations for RP of this family.

Objective: To investigate the clinical efficacy and safety of adefovir dipivoxil (ADV) in combination with bicyclol for the treatment of chronic hepatitis B (CHB) in seniors., Methods: 96 senior patients with CHB were randomly divided into two groups, the treatment group and the control group. On the basis of routine liver protective treatment, patients in the treatment group received ADV (10 mg/d) and bicyclol tablets (25 mg, tid.) orally, and those in the control group were orally administrated ADV tablets (10 mg/d) only. The treatment course for both groups was 24 weeks. Serum ALT, AST, and alterations of virological parameters were observed before and after the treatment., Results: Before and at the end of the 24 weeks treatment, ALT level for the treatment group was (208.44 +/- 94.22) and (34.47 +/- 12.79) U/L, and those for the control group was (205.73 +/- 96.48) and (44.20 +/- 21.96) U/L, respectively (difference between groups P < 0.01). At the end of the 24 weeks treatment, ALT normalization rates for the treatment group and the control group were 76.6% and 54.5%, respectively, and AST normalization rates for them were 76.6% and 54.5%, respectively (both differences between groups P < 0.05); HBV DNA loads for the treatment group and the control group were decreased by (3.1 +/- 1.40) lgIU/ml and (2.98 +/- 1.17) lgIU/ ml, respectively (difference between groups P > 0.05). The incidence rates of adverse events between two groups were not statistically significant., Conclusion: It suggested that the treatment of ADV in combination with bicyclol for senior patients with CHB is effective and safe.

Nature-approximating landscape architecture (NALA) is a concept of sustainable development as applied to landscape architecture, while the urban habitants' awareness and acceptance of NALA idea is the key for the successful application of NALA. Through semi-structured interview, this paper explored the attitudes of the habitants in Hongshan District of Wuhan City toward the NALA design and management, and the influence of the social-economic characteristics of the responders on their attitudes toward the NALA. A fairly low percentage of the responders approved of the NALA design (10.3% - 46.9%) and management (7.4% - 34.9%). The attitudes towards NALA design were mainly affected by the responders' age, and the attitudes toward NALA management were significantly correlated with the responders' age, educational level, and profession. The efficient cause why a large number of responders did not support the NALA was that these responders attached importance to the aesthetic effect of green space, and preferred cleanliness and order. The lack of related ecological knowledge and environmental awareness was the root cause of the lesser support towards NALA. To establish NALA demonstration bases and to intensify the publicity and education of NALA idea and related ecological knowledge could promote an increasing number of urban habitants actively participating in NALA construction.

Aim: To develop a coxsackievirus A16 (Cox A16) VP1 gene plasmid which delivered by live attenuated Salmonella., Methods: The plasmid which expressed VP1 protein of CoxA16 was constructed by gene recombination. Cellular expression was assessed by Western bloten analysis. Then the recombinant attenuated Salmonella which harboring the plasmid were constructed by electro transformation., Results: CoxA16 VP1 gene sequence was inserted into a eukaryotic expression plasmid. VP1 protein was detected in the culture supernatant., Conclusion: The plasmid is constructed successfully and it can be expressed effectively in vitro. The recombinant bacteria are constructed successfully. This has provided a basis for further research of an oral CoxA16 vaccine.

Objective: To develop attenuated Salmonella which harboring enterovirus 71 (EV71) VP1 gene., Methods: The plasmid which expressed VP1 protein of EV71 was constructed by gene recombination. Cellular expression was assessed by Western Blot analysis. The recombinant plasmid was then transformed into attenuated Salmonella SL7207., Results: EV71 VP1 gene sequence was inserted into a eukaryotic expression plasmid VR1012. VP1 protein was detected by Western Blot analysis in the culture supernatant. And the attenuated Salmonella harbored the plasmid stable., Conclusion: The plasmid was constructed successfully and it can express effectively in vitro. The bacteria which harboring the plasmid were constructed successfully. This has provided a basis for further research of an oral EV71 vaccine.

Aim: To analyze mutations in transforming growth factor beta-induced (TGFBI) gene in a Chinese pedigree with Reis-Bücklers corneal dystrophy (RBCD, also known as GCD3)., Methods: In a five-generation Chinese family, eight members were identified with RBCD and the rest were unaffected. All members of the family underwent complete ophthalmologic examinations. Exons of TGFBI were amplified by polymerase chain reaction, sequenced, and compared with a reference database., Results: A single heterozygous C>T (R124C) point mutation was found in exon 4 of TGFBI in all the affected members of the pedigree, but not in the unaffected members., Conclusion: R124C which was a known mutation for lattice corneal dystrophy type I, segregated with the RBCD in this pedigree. This elucidated the correlation between genotype and phenotype in a Chinese family of RBCD.

Purpose: Familial nystagmus complicated with cataract and iris anomalies are genetically heterogeneous, and the pathophysiological mechanisms remain unclear. It is anticipated that mutations in the paired box 6 (PAX6) gene play a major role in pathogenesis of malformations in anterior segment of the eye. In this study, we analyzed PAX6 in a Chinese pedigree of nystagmus, cataract and iris anomalies. This study will provide insights into the genetic basis of this disease., Methods: Complete ophthalmologic examinations were performed on four patients (excluding one dead patient) and four unaffected individuals in this four-generation family. All coding exons of PAX6 were amplified by polymerase chain reaction (PCR), sequenced and compared with reference database. The variations detected were evaluated in available family members as well as 110 normal controls. Possible changes in structure and function of the protein induced by amino acid variance were predicted by bioinformatics analysis., Results: Nystagmus, cataract or iris anomalies were found in all patients of this family, but the severity was different among these patients. A novel missense mutation in PAX6 was identified in all affected individuals but not in asymptomatic members and 110 normal controls. This mutation causes an amino acid substitution of proline to glutamine at position 118 (p.P118Q) of the paired domain of the PAX6 protein. Such a change may cause structural and functional changes of the protein based on bioinformatics analysis., Conclusions: This study added a novel mutation to the existing spectrum of PAX6 mutations, suggesting that a mutation in PAX6 correlated with anterior segment disorders observed in this family.

Purpose: To analyze two candidate genes, trabecular meshwork inducible glucocorticoid response (MYOC/TIGR) and human dioxin-inducible cytochrome P450 (CYP1B1), in a Chinese pedigree of primary open-angle glaucoma., Methods: In a three-generation family containing 14 members, four of them were patients with primary open-angle glaucoma, one was a glaucoma suspect, and the rest were asymptomatic. All members of the family underwent complete ophthalmologic examinations. Exons of MYOC and CYP1B1 were amplified by polymerase chain reaction, sequenced, and compared with a reference database., Results: Elevated intraocular pressure and impaired visual field were found in all patients. One MYOC heterozygous mutation G367R, in exon 3 was identified in four patients and the suspect, but not in the rest of the family members. Meanwhile, four single nucleotide polymorphisms in MYOC and CYP1B1 genes were found., Conclusions: Although the G367R mutation of MYOC, which causes primary open-angle glaucoma in the form of autosomal dominant inheritance, has been reported in some other ethnicities, it was found in Chinese pedigree for the first time.

Aim: To analyze phenotype and genotype of a Chinese pedigree with Avellino corneal dystrophy (ACD)., Methods: Complete ophthalmic examinations were performed on all the family members. Exons of TGFBI were amplified by polymerase chain reaction, sequenced, and compared with a reference database., Results: A single heterozygous G>A (R124H) point mutation was identified in exon 4 of TGFBI in three affected members and two unaffected children who were offsprings of the affected members, but not in the other family members., Conclusion: Mutation R124H in TGFBI was identified in this pedigree and appeared to be the disease causing mutation. Atypical phenotype and low penetrance was observed in this pedigree.

Aim: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells., Methods: HTM cells were cultured to confluent and incubated with 0.4µmol/L Dex and/or 0.05µmol/L LAT-A. FN expression in HTM cells was evaluated by Western blot and immunofluorescence microscopy., Results: Dex up-regulated FN production in HTM cells, failed to do so when co-incubated with LAT-A. LAT-A decreased production of FN in cultured HTM cells., Conclusion: This study indicated that LAT-A may modulate the expression of fibronectin in trabecular meshwork to achieve treatment for steroids and other types of glaucoma. It has an important prospect as an intraocular pressure- lowering drug.

Purpose: To characterize the effects of circadian rhythm, feeding time, age, general anesthesia, and ocular hypotensive compounds on intraocular pressure (IOP) of the Tibetan monkey (Macaca thibetana)., Methods: Tibetan monkeys were trained for IOP measurement with the TonoVet® rebound tonometer without sedation or anesthesia. Their circadian IOP fluctuation was monitored every 3 h. Effects of changing the feeding time, general anesthesia, age (2-3 year-old versus 8-15 year-old animals), and various pharmacological agents, such as travoprost, timolol, naphazoline and spiradoline, on IOP were also evaluated., Results: After behavioral training, conscious Tibetan monkeys were receptive to IOP measurement. The lowest and highest IOP values in a circadian cycle were recorded at 3:00 AM (19.8±0.4 mmHg, mean±SEM, n=12) and noon (29.3±0.9 mmHg), respectively. Changing the feeding time from 11:30 AM to 12:30 PM lowered the noon IOP to 25.1±1.2 mmHg. General anesthesia lowered IOP in these monkeys, while IOP of young and mature animals were similar. Three hours after topical ocular administration, travoprost reduced IOP by 5.2±0.6 mmHg (n=6, p<0.001), and timolol reduced IOP by 2.8±0.7 mmHg (p<0.05). Naphazoline and spiradoline lowered IOP by 4.8 mmHg and 2.5 mmHg (both p<0.001), respectively, 2 h after drug administration., Conclusions: The circadian IOP fluctuation in conscious Tibetan monkeys and their responses to travoprost, timolol, and other experimental conditions are similar to other primates. These monkeys appear to be a suitable model for glaucoma research.

Objective: To explore the clinical significance of prenatal ultrasonic diagnosis of fetal nephrohydrosis and its prognosis., Methods: Prenatal ultrasonography was performed on 9526 women at more than 20 weeks gestation, and 162 women whose anteroposterior diameter of the renal pelvis was > or = 8 mm were included in this study. The grade of fetal nephrohydrosis was classified according to Grignon grading method. The changes in fetal nephrohydrosis were observed regularly until delivery., Results: (1) The incidence of fetal encephalic fluid and Grignon grade: there were 162 fetuses with fetal nephrohydrosis among 9526 fetuses, with an incidence of 1.7%. The numbers of fetuses with Grades 1 to 5 were 71, 59, 7, 3 and 22, respectively. (2) The distribution in gestational weeks of fetal nephrohydrosis: generally, fetal nephrohydrosis was first diagnosed at (33 +/- 5) weeks, and the maximum degree of nephrohydrosis was observed at (36 +/- 3) weeks. One hundred and ten fetuses with nephrohydrosis recovered during the process of gestation, at about (37 +/- 4) weeks. (3) Poor terminations of pregnancy: there were 40 (25%) fetuses with poor terminations among totally 162 cases. Among these 40 fetuses, 3 (27%) were first diagnosed at 20 to 24 weeks within 11 fetuses, 6 (24%) were first diagnosed at 25 to 28 weeks within 20 fetuses, 14 (26%) were first diagnosed at 29 to 32 weeks within 53 fetuses, 11 (23%) were first diagnosed at 33 to 36 weeks within 48 fetuses, and 6 (20%) were first diagnosed at 37 to 40 weeks within 30 fetuses. The results demonstrate that high grade of fetal nephrohydrosis according to Grignon grading method indicates a poorer prognosis of the fetus. (4) Follow-up results: there were 122 (75%, 122/162) live and healthy neonates in total, including 110 neonates whose nephrohydrosis recovered prior to birth and 12 neonates whose nephrohydrosis recovered within 1 week after birth. These neonates all developed well without any urinary sequela within the first 2 years. There were also 20 (12%, 20/162) hydronephrotic neonates who were still diagnosed as nephrohydrosis within 1 week after birth, including 11 fetuses graded below Grignon grade 3, whose nephrohydrosis recovered 3 to 12 months after birth and who developed well without any urinary symptoms., Conclusions: The study demonstrates that hydronephrotic fetuses who are diagnosed at earlier gestational weeks and with higher grading have poorer prognosis. The Grignon grading method can be used in the prenatal evaluation of fetal nephrohydrosis to predict the prognosis of the fetus.

Background: Although angiogenesis and lymphangiogenesis in gastrointestinal cancers has been investigated in many studies, their distribution characteristics in gastrointestinal intramucosal tumors have not been well addressed., Methods: We evaluated the blood microvessel density (BMVD) and lymphatic microvessel density (LMVD) by immunostaining with monoclonal antibodies of CD34 and D2-40 in 37 patients with stomach intramucosal carcinoma and 28 patients with colorectal intramucosal neoplasia. Microvessels with endothelial cells labeled by CD34 but not by D2-40 were recognized as blood microvessels; and microvessels with endothelial cells labeled by both CD34 and D2-40 were recognized as lymphatic vessels. Furthermore, the relationships between expression of vascular endothelial growth factor (VEGF), VEGF-C, and BMVD, LMVD were investigated as well., Results: The LMVD was significantly higher in peritumoral tissues than in corresponding normal tissues in gastrointestinal intramucosal tumors (20.87 versus 14.56, P = 0.003). However, there was no significant difference in the BMVD between peritumoral tissues and corresponding normal tissues (P = 0.166). The BMVD in peritumoral tissues was higher in patients with lymph node metastases than in patients without lymph nodes metastases (P = 0.047). Our results did not show significant association between VEGF, VEGF-C and BMVD, LMVD., Conclusions: Our results suggested that the increase of lymphangiogenesis seems superior to the increase of angiogenesis in gastrointestinal intramucosal tumors.

Aim: To explore the influence of GABAergic neurotransmitters and GABAA receptors on the auditory afferent impulses recorded in the brainstem evoked by electro-stimulation., Methods: Brainstem slices were prepared using ddy/ddy mice of postnatal 0-5th days. The brainstem slices were stained with a voltage-sensitive dye(NK3041). The cut end of the vestibulocochlear nerve (nVIIIth) connected with slices was stimulated by a tungsten electrode, a 16 x 16 pixels silicon photodiode array apparatus was used to record the optical mapping from auditory brainstem slices. The data were analyzed by ARGUS-50/PDA software., Results: The spatial-temporal patterns of the excitatory propagation from the vestibulocochlear nerve (nVIIIth) to cochlear nucleus and vestibular nucleus were displayed with multiple-sites optical recording. The optical signal coming from one pixel consisted of a fast spike-like response and a following slow response. Inhibitory neurotransmitter GABA decreased the fast spike-like response and following slow response of evoked optical signals, while an antagonist BMI against GABAA receptors increased the both responses., Conclusion: A 16 x 16 pixel silicon photodiode array apparatus can be used to record multiple-sites optical mapping evoked by electro-stimulation to the cut end of the vestibulocochlear nerve. The every optical signal consists of both presynaptic and postsynaptic elements. Inhibitory neurotransmitter GABA and an antagonist BMI of GABAA receptors can modulate the excitatory propagation of evoked optical signals.

Objective: To investigate clinical features of the patients with hepatitis B superinfected with acute hepatitis E (AHE)., Methods: Totally 625 consecutive patients enrolled from Dec 2002 to Dec 2006 were studied retrospectively. All of the patients were subclassified into acute hepatitis E group (AHE=437 cases) and Superinfected Group (S=188 cases), and S group was further divided into the group of chronic hepatitis B superinfected with acute hepatitis E (CHB+AHE, 130 cases) and the group of liver cirrhosis and hepatitis B superinfected with acute hepatitis E (LCB+AHE, 58 cases). In 32 of the 188 superinfected patients the effects of HEV on HBV were observed by comparing the levels of HBV DNA in acute vs. convalescence stages., Results: Compared with the patients with AHE, the superinfected patients had a higher level of total bilirubin (TBil), an elevated frequency of fulminate hepatitis, mortality and a longer period of the mean hospital stay for the cured patients but significantly lower levels of alanine aminotransferase (ALT), serum albumin and prothrombin activity (PA). Furthermore, the group of LCB+AHE had a higher level of TBil and higher incidences of complications such as ascites, peritonitis, hepatic encephalopathy and disturbance in glycometabolism than the group of CHB+AHE. The follow-up for the superinfected patients showed that 20 of 32 patients (62.5 percent) had decreased copies of HBV DNA during the recovery phase compared with the acute phase, and the mean decrease of HBV DNA was 2.1 log10. The HBV DNA was in a persistently undetectable level in 6 of 32 (18.8 percent) superinfected patients. However, 4 of 32 patients (12.5 percent) showed an unchanged levels of HBV DNA and 2 cases (6.2 percent) had a slightly increased HBV DNA levels., Conclusion: Superinfection with AHE in patients with chronic hepatitis B leads to a more severe hepatic damage and the replication of HBV DNA can be transiently inhibited.

Objective: To explore the diagnosis, clinical course and prognosis of fetal multicystic kidney dysplasia (MCDK)., Methods: 24 858 pregnant women detected by prenatal ultrasound, here were 41 cases with fetal multicystic kidney dysplasia, these fetuses were diagnosed at average 29.8 weeks of gestation, Carried on an observation to fetuses with multicystic kidney dysplasia and postnatal follow-up study., Results: T17 cases were induced abortion. Of 13 infants, 1 case involute, 3 cases decrease, 9 cases no change., Conclusion: Prenatal ultrasonography can actual diagnosis for fetal multicystic kidney dysplasia, the key of management of multicystic kidney dysplasia is assessment of fetal prognosis, the natural history of unilateral MCDK is usually benign, the affected kidneys tend to show involution after birth. But bilateral MCDK often associated with impairement of renal function, abnormal chromosome or other anomalies, which indicates a poor prognosis.

Objective: To discuss the clinical significance of fetal encephalic accumulated fluid revealed by prenatal ultrasonography., Methods: Prenatal ultrasonography was performed on 8426 women at more than 20 weeks' gestation. Totally 150 women with fetal encephalic accumulated fluid more than 5 mm were included in this study. The changes of fetal encephalic accumulated fluid and the associated anomalies were observed regularly every 2 weeks until delivery. The live infants were followed up regularly., Results: The incidence of fetal encephalic fluid was 1.8%, including 72 cases with fluid in the fetal anterior or posterior cornu of unilateral ventricle, 46 cases with accumulated fluid in fetal posterior fossa, 32 cases with fluid in more than 2 sites. Generally, the accumulated fluid in fetal encephalus was first diagnosed at 17 - 40 gestational weeks, with a median of (26 +/- 5) weeks. Most of them were found between 29 - 32 gestational weeks (63 cases, 42.0%), and the maximum amount of accumulated fluid was also found between 29 - 32 weeks (70 cases, 46.7%). Spontaneous regression of intracranial fluid could be seen in 111 fetuses (74.0%). The period of fluid regression ranged from 29 to 40 weeks of gestation, of which the average gestational week was (36 +/- 2) weeks. Additionally, the most frequent period of regression was in the first two thirds of the three trimesters of pregnancy. The incidence of defected infants was 3.8%, 10.2% and 67.4%, respectively, when the amount of accumulated fluid was less than 10 mm, 10 - 14 mm and more than 15 mm. And the accumulated fluid in more than 2 sites was also a risk factor of defected fetuses, with an incidence of 60.0%., Conclusions: Most cases could be diagnosed between 29 - 32 gestational weeks, and the maximum amount of accumulated fluid is also observed in this period. The more fluid in fetal encephalus, the more sites the fluid distributed in, the more defected fetuses or infants would be observed. So in cases of more than 15 mm of fluid, or accumulated fluid in more than 2 sites, anomalies should be observed extremely carefully.

Objective: To investigate the expression of osteopontin mRNA and its correlation with clinicopathologic features of gastric cancer and elucidate its role in tumor invasion and distant metastasis., Methods: The expression of OPN mRNA was detected by semi-quantitive RT-PCR. The relationship between the relative content of OPN mRNA and clinicopathologic features of gastric cancer was analyzed., Results: In 66 cancer tissue samples, a 330 bp band was detected in 50 cases, the positive rate of OPN mRNA expression was 75.8% (50/66). The expression in all 20 cases of normal gastric mucosa was negative. The expression was associated with the depth of tumor invasion, diameter, lymph node metastasis and but had no correlation with differentiation grades. The 66 patients were followed up for 10 approximately 27 months (mean 16 months). The OPN mRNA expression positive group (50 cases) had recurrence in 15 patients and the negative group (16 cases) had only 1 case with recurrence (P = 0.05); 10 patients died in OPN mRNA expression positive group but no patient died in OPN staining negative group (P = 0.05)., Conclusion: OPN mRNA is over-expressed in gastric cancer. It reflects the progression of disease and association with poor prognosis of gastric cancer. OPN may play an important role in the process of distant metastasis in gastric cancer.

Objective: To explore the risk of developing malignant neoplasm in a cohort with the history of environmental exposure to crocidolite asbestos., Methods: A retrospective cohort and follow-up study covering 15 years (1987 --> 1995 --> 2001) was carrid out in a small town in Yunnan province. The cohort comprised 6254 local inhabitants. The deaths from and the RR of asbestos-related malignant neoplasms were studied., Results: There were 186 deaths from neoplasms in the observation group, the mortality being 2160.5 per 10(6) person-years (RR=1.293, 95%CI: 1.032-1.618), 20 of those deaths were caused by mesothelioma, with a crude mortality of 232.3 per 10(6) person-years (RR=17.929; 95%CI: 2.406-133.592). The mortalities for men and women were 267.5 and 186.7 per 10(6) person-years respectively. The crude mortality from lung cancer (56 deaths) was 650.5 per 10(6) person-years,there is no significance between the two groups (RR=1.434; 95%CI: 0.968-2.486). The difference in mortality from gastrointestinal cancer between the two groups is not significant, whereas the risk of man intestinal cancer in the observation is significant (RR=3.71; 95%CI: 1.077-13.270)., Conclusion: The risk of developing mesothelioma is significantly increased in the population with environmental exposure to crocidolite. The risk of man intestinal cancer in the town awaits additional studies.

gamma-Aminobutyric acid (GABA)ergic neurons play important tropic and modulatory roles in the auditory pathway, especially in the early stage between postnatal Days 0 and 5. The effects of GABA and GABAa receptor antagonist were observed in this experimental study. Numerous histological and electrophysiological studies have been performed on the contribution of GABA to the auditory pathway; however, the spatio-temporal patterns of excitatory propagation and the relationships between GABA receptor and excitatory propagation have yet to be reported. Using an optical recording technique and a voltage-sensitive dye, the spatio-temporal patterns of excitatory propagation were observed in the auditory brainstem slices of early postnatal mice. A bath containing 50 microM GABA was applied, which largely inhibited the excitatory activities along the vestibulocochlear pathway. Bicuculline methiodide (BMI), a competitive antagonist against GABAa receptor, partially reversed the effects of GABA on the optical signals. Bath application of BMI alone helped to facilitate the depolarization course and its effect was apparent as an enlargement of the depolarized region from the cochlear nucleus and vestibular nucleus to some adjacent brainstem nuclei, as well as enhancing the amplitude of changes in the optical signals. The experimental results seem to suggest that GABAa receptors are widely distributed in an early postnatal auditory brainstem. GABA exhibited a greater modulating effect in the adjacent brainstem nuclei, which are involved in complex information processes, than that observed in the modulating primary auditory pathway. In the present experiment, significant GABAergic contributions to the optical recordings in the auditory brainstem were observed.

Background: To investigate the effect of Oxymatrine (OM) on serum cholinesterase (ChE) during the treatment of viral hepatitis and the relationship between the change of ChE and the change of albumin (ALB), prothrombin activity (PTA) and other liver function tests., Methods: A total of 98 patients with viral hepatitis were divided into four groups. Group A consisted of 31 patients and were treated with OM intravenous infusion; Group B consisted of 30 patients, treated with OM orally; Group C consisted of 7 patients and were treated with OM intramuscular injection while Group D consisted of 30 patients, and were not treated with OM. ChE, ALB, PTA, liver function, renal function, soluble complement receptor-1 (sCR1) and erythrocyte innate immune adhesion function (EIIAF) were regularly determined., Results: ChE in Group A,B,C was dropped obviously during the treatment (P less than 0.001, less than 0.001, 0.023=. But there were no change in ALB, PTA, sCR1, EIIAF (P greater than 0.05), and remarkable improvement of ALT, AST, TBiL was seen during the treatment in Groups A, B, C. After the treatment with OM, the level of ChE recovered soon., Conclusion: Serum level of ChE significantly declined during the treatment of viral hepatitis with OM, but no change was found in ALB, PTA, sCR1, EIIAF while liver function tests showed better results. So the drop of ChE does not mean deprivation of patient's liver disease.

Aim: To investigate the clinical pathologic features of gastrointestinal leiomyoma and the diagnostic value of endoscopic ultrasonography (EUS) on gastrointestinal leiomyoma., Methods: A total of 106 patients with gastrointestinal leiomyoma diagnosed with EUS were studied. The location, size and layer origin of gastric and esophageal leiomyomas were analyzed and compared. The histological diagnosis of the resected specimens by endoscopy or surgery in some patients was compared with their results of EUS., Results: The majority of esophageal leiomyomas were located in the middle and lower part of the esophagus and their size was smaller than 1.0 cm, and 62.1 % of esophageal leiomyomas originated from the muscularis mucosae. Most of the gastric leiomyomas were located in the body and fundus of the stomach with a size of 1-2 cm. Almost all gastric leiomyomas (94.2 %) originated from the muscularis propria. The postoperative histological results of 54 patients treated by endoscopic resection or surgical excision were completely consistent with the preoperative diagnosis of EUS, and the diagnostic specificity of EUS to gastrointestinal leiomyoma was 94.7 %., Conclusion: The size and layer origin of esophageal leiomyomas are different from that of gastric leiomyomas. Being safe and accurate, EUS is the best method not only for gastrointestinal leiomyoma diagnosis but also for the follow-up of patients.

Objective: To determine the causes of choledochal dilatation in patients with obstructive jaundice., Methods: One hundred and sixty-four patients with obstructive jaundice were investigated by endoscopic retrograde cholangiopancreatography(ERCP), and patients with choledochal dilatation (group I, n=110) were compared with those without choledochal dilatation (group II, n=54)., Results: The causes of common bile duct dilatation were choledocholith, juxtapapillary duodenal diverticula and congenital dilatation of the common bile duct. The distal common bile duct and its surroundings were abnormal in 104 (94.55%) of the 110 patients and in 13 (24.08%) of the 54 patients (P<0.01). Juxtapapillary duodenal diverticulum accounted for 24.55% in group I, and only in 9.26% in group II (P<0.05). Post-cholecystectomy patients were 13.64% in group I, and only 5.56% in group II., Conclusions: The abnormalities of the distal common bile duct and its surroundings can usually be detected as underlying causes of common bile duct dilatation. ERCP is necessary before cholecystectomy, since it is considered the "gold standard" for the diagnosis of distal common bile duct abnormalities.

Aim: To evaluate the role of ELAM-1 and ICAM-1 in the course of inflammatory reactions during focal brain ischemia/reperfusion., Methods: The focal brain ischemia/reperfusion model is carried by occluding middle cerebral artery. The expression of ELAM-1 and ICAM-1 mRNA after ischemia/reperfusion was evaluated with RT-PCR., Results: No ELAM-1 and ICAM-1 mRNA were detected in the sham-operated cortex and only little in the nonischemic cortex. The expression of ELAM-1 and ICAM-1 mRNA were upregulated at 1 hour, peaked at 6 hour and 3 hour respectively and remained elevated for up to 48 hours after ischemia/reperfusion., Conclusion: ELAM-1 and ICAM-1 participate in brain injury during focal ischemia/reperfusion and both of them play an important role in leukocyte infiltration into the ischemic tissues.

The beta-thalassemia syndromes are a heterogeneous group of genetic disorders characterized by reduced or absent expression of the beta-globin gene. To date, over 300 beta-thalassemia alleles have been characterized in or around the beta-globin region. Thalassemia major is severe anemia necessitating chronic blood transfusions, splenectomy, iron chelation therapy, and bone marrow transplantation. Usually thalassemia major results from homozygosity or compound heterozygosity for severe betaO- and/or beta+-thalassemia mutations. Thalassemia intermedia is a clinical diagnosis that describes a symptomatic but less severe condition than beta-thalassemia major. beta-thalassemia intermedia may arise from several different combinations of alpha- and/or beta-thalassemia mutations. Heterozygous beta-thalassemia is typically characterized by a mild microcytic hypochromic anemia without any significant clinical implications. In this report, we describe a 63-year-old Africian American woman with asymptomatic homozygous beta-thalassemia, who seems to carry 2 copies of the -29 mutation in the promoter region of the beta-globin gene. Her elevated hemoglobin F level of 83% was associated with heterozygosity for the Xmn I polymorphism upstream of the Ggamma-globin gene. Southern blot analysis at the alpha-globin locus did not show any deletion that would account for the mildness of her phenotype. Therefore, homozygosity for the -29 mutation along with the Xmn I polymorphism appears to confer an extremely mild beta-thalassemia phenotype. This observation has important implications in the prenatal diagnosis and genetic counseling of families segregating this type of genetic defect.

While analyzing 280 hemoglobinopathy kindreds with prescribed molecular tests, 3 unusual mutations were observed that required additional characterization. In the first case, the hypervariable region flanking the alpha-globin genes generated an intermediate length 8.2 kb psi zeta-globin gene fragment on a Southeast Asian chromosome with two deleted alpha-globin genes. Rehybridization of the Southern blot with alpha-globin probe distinguished the mutation unambiguously. In the second case, restriction enzyme analysis of a PCR amplified black beta-globin gene detected a novel beta-83 point mutation adjacent to a promoter element. In the third case, which was uninformative with available allele specific oligonucleotides (ASOs), total genomic PCR amplification and sequencing identified a single basepair insertion in codon 36/37 of an Iranian beta-globin gene that shifted the reading frame and obliterated gene activity. Developing additional region-specific ASOs will further diminish the number of cases that must be characterized by genomic PCR sequencing.