Identification of genotype of chlamydia trachomatis from outpatients of trachoma / 中华实验眼科杂志

Background Trachoma is a common disease in ophthalmology,but few reports about the pathogenetic genotype are reported.Objective This study was to investigate the genotypes of Chlamydia trachomatis.Methods Conjunctival specimens were collected in 16 patients with trachoma in Beijing Tongren Eye Center from January,2003 and August,2006.Variable sequence 4 (VS4) of ompl fragment was amplified by nested PCR(n-PCR) using specific primer of Chlamydia trachomatis,and then the PCR products were sequenced.The DNA sequences were analyzed by software Clustal X and MEGA2,and the genetic characteristics were compared with the known sequences of GenBank.Results The PCR product fragment of MOMP gene of trachoma was 277 bp.Three types of Chlamydia trachomatis strains were idcntified in the 16 specimens,including B-genotype in 9 strains (56.3％),C-genotype in 4 strains(25.0％) and D-genotype in 3 strains(18.7％).High homology was seen in the gene sequences of Chlamydia trachomatis strains between the B-genotype or C-genotype strains and the same genotypes of GenBank or those from some districts of China.However,some differences were exhibited among the Dgenotype strains.Conclusions Identification of genotype of Chlamydia trachomatis has differential effect on trachoma and inclusion conjunctivitis.

Cross-linking of human amniotic membrane by ultraviolet A-riboflavin / 中华实验眼科杂志

BackgroundStudies confirmed that ultraviolet A (UVA)- riboflavin photodynamic therapy can control keratoconus progresses by altering the physicochemical property of cornea.The collagen components of amniotic membrane transplantation is similar to that of cornea and amniotic membrane transplantation has been widely used to ocular surface reconstruction.However,the study on UVA riboflavin-induced-collagen crosslinking for amniotic tissue is less now.ObjectiveThis study was to investigate the role of UVA-riboflavin on frozen-preserved human amniotic membrane.Methods Human amnions were obtained in informed consent and prepared into 2 mm×15 mm pieces and were then divided into 4 groups using lottery method and 6 pieces for each group.The first 3 groups were treated with the photosensitizer riboflavin and UVA-irradiation ( wavelength:370 nm ; irradiation energy:1,2 or 3 mW/cm2,distance:10 mm) for 30 minutes,and the untreated fourth group was as control group.Biomechanical stress-strain test was performed using a microcomputer-controlled biomaterial tester and the stress(mN) was recorded when the strains were set to 5％,10％ and 15％.The 7 mm diameter of human amniotic membrane pieces were trephined and divided into 4 groups(5 pieces for each group) with the treated method as mentioned above,and then the buttons were exposed to 0.1％ collagenase Ⅰ solution.The transparency was scored and the complete dissolving time was record.In histological evaluation,three groups (3 pieces for each group) of human amniotic membranes were treated using UVAriboflavin(3 mW/cm2),0.1％riboflavin,normal saline for 30 minutes respectively and examined under the transmission electron microscopy.This study was performed under the permission of the Ethic Commission of Beijing Tongren Hospital.ResultsWhenthestrainwas 5％,10％,15％,thestressof controlgroupand1,2,3 mW/cm2UVA group were statistically signifcantly different ( F =3.411,P =0.037; F =9.927,P =0.001;F=11.118,P=0.000).The tensile strength of human amniotic membrane cross-linked with UVA-riboflavin was statistically significantly increased in comparison to the control group (P＜0.05 ),and the tensile strength of human amniotic membrane became stronger as UVA power increased.The complete dissolve time was (8.6± 1.8 ) hours for the control group,(39.6± 2.3 ) hours for 1 mW/cm2 UVA group,(71.4±0.9 ) hours for 2 mW/cm2 UVA group,(78.8± 1.8 ) hours for 3 mW/cm2 UVA group,showing the enhanced anti-enzyme ability of human amniotic membrane after cross-linking(P＜0.01 ).The collagen density in the UVA-riboflavin treated group was increased,the connection among the collagen fibers as well as between the stroma and the epithelium became tighter than those of control group.ConclusionsCollagen cross-linking with UVA-riboflavin make the biomechanical strength and enzymatic resistance of human amniotic membrane enhance and ultrastructure change of human amniotic membrane.

Diagnosis of 22q11 deletion and duplication in congenital heart disease by multiplex ligation dependent probe amplification / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical utility of multiplex ligation-dependent probe amplification (MLPA) for detecting 22q11 deletion and duplication in congenital heart disease (CHD) cases and to study the incidence of 22q11 deletion and duplicaton in different kinds of CHD.</p><p><b>METHODS</b>Forty eight probes of which 25 located in 22q11 low copy number region (LCR 22s A-H), 7 in 22q11 surrounding region (CES, 22q13) and 16 in chromosomes 4, 8, 10 and 17 were selected to detect 22q11 deletion and duplication in 181 preoperative children with CHD and 14 fetuses with serious CHD or CHD with multiple malformations. In these cases, karyotype analysis was also performed.</p><p><b>RESULTS</b>MLPA demonstrated that 7 cases had 22q11 deletion [6 cases from CLTCL1 to LZTR1(LCR A-D) and 1 case from CLTCL1 to PCQAP (LCR A-C)] and that 1 case had 22q11 duplication,spanning from ZNF74 to LZTR1(LCR B-D). The phenotypes of heart defect included ventricular septal defect, atrioventricular septal defect, pulmonary stenosis and tetralogy of Fallot. Karyotype analysis showed that 1 case had 21q deletion [46, XY, 21q], 1 case had mosaic trisomy 8 [47,XY, +8/46, XY(1:2)] and 4 cases had trisomy 21. One of the 4 cases with trisomy 21 had concurrent 22q11 duplication.</p><p><b>CONCLUSIONS</b>MLPA is a rapid, sensitive, site specific and relatively inexpensive method for diagnosis of 22q11 deletion and duplication in CHD. 22q11 deletion and duplication may cause various kinds of CHD, suggesting that genetic detection should be performed routinely in CHD patients.</p>

Loss of imprinting of IGF2 in cord blood of newborns of Chinese Han population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the imprinting status of IGF2 and phenotypes of loss of imprinting (LOI) in cord blood of neonates of Chinese Han population and to investigate relative factors to LOI.</p><p><b>METHODS</b>Cord blood of 1010 Chinese Han newborns were collected and the imprinting status of IGF2 was detected by reverse transcription-PCR(RT-PCR) and restriction fragment length polymorphism.The relationships between LOI and fetal growth indices, features of parents and grandparents, clinical characteristics were analyzed.</p><p><b>RESULTS</b>Of all cases, 42.8% (432/1010) were heterozygous for a polymorphism of Apa I site in exon 9 of IGF2, while 21.6%(66/306) displayed IGF2 LOI. Maternal factors including average age, gestational age, BMI pre-pregnancy, weight gain during pregnancy and the level of HB, HCT, and other indices of biochemistry in their second and third trimester were not correlated with LOI expression. However in newborns with fathers older than 35 yrs, 31.7%(19/60) displayed LOI, which was significantly more common than that in newborns with younger fathers (P< 0.05, chi square is 4.69). There were no difference in birth weight (BW) between normal imprinting and LOI groups. But if the newborn's weights were in 2500-2999 g, LOI was 6.25%(2/32), which was significantly lower than that in 3000 g group (P< 0.05, chi square is 4.89). In groups with BW being less than 2500 g and more than/equal to 4000 g, the LOI newborn's blood glucose was decreased significantly after 2 hrs (P< 0.01, t is 7.47 and 10.9).</p><p><b>CONCLUSION</b>In newborns of Chinese Han population, 21.6% showed IGF2 LOI in cord blood. IGF2 LOI may have some influences on fetal growth. Paternal age is associated with LOI.</p>

Etiological analysis on ocular fungal infection in the period of 1989 - 2000 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>This study was to review the distribution and shifting trend of fungal of culture specimens isolated from eyes of patients at the Beijing Institute of Ophthalmology, Tongren Hospital, Beijing, China.</p><p><b>METHODS</b>The fungal culture-positive rate, the distribution and change of isolates of 2609 specimens collected in a 12-year period (1989 - 2000) were retrospectively analyzed.</p><p><b>RESULTS</b>In 775 positive cultures, 707 specimens (91.2%) were from the cornea, 22 (2.8%) from the conjunctiva, 15 (1.9%) from the anterior chamber, 9 (1.2%) from the vitreous body, 3 (0.4%) from the lacrimal sac, and 19 (2.5%) from other parts of the eye. The average culture-positive rate was 29.7%. The ratio of the positive cultures in the first half year (from January to June) to those in the second half (from July to December) was 1:2.1. The main genus cultured was Fusarium sp (58.7%), followed by Aspergirum sp (16.8%). The percentage of Fusarium sp was increased from 53.6% (1989 - 1994) to 60.2% (1995 - 2000), whereas the percentage of Aspergirum sp was decreased from 22.3% (1989 - 1994) to 15.1% (1995 - 2000).</p><p><b>CONCLUSIONS</b>Fusaruim sp is one of the most predominant pathogens of ocular fungal infection in northern China and its incidence tends to increase, but that of Aspergirum sp to decrease. It is very important to recognize the distribution and shifting trend of pathogenic fungi in the diagnosis, prevention and treatment of fungal keratitis.</p>

Analysis of human transforming growth factor beta-induced gene mutation in corneal dystrophy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Corneal dystrophy is a group of inherited blinding diseases of the cornea. This study was to identify the mutations of the keratoepithelin (KE) gene for proper diagnosis of corneal dystrophy.</p><p><b>METHODS</b>Three families with corneal dystrophy were analysed. Thirteen individuals at risk for corneal dystrophy in family A, the proband and her son in family B, and the proband in family C were examined after their blood samples were obtained. Mutation screening of human transforming growth factor beta-induced gene (BIGH3 gene) was performed.</p><p><b>RESULTS</b>Five individuals in family A were found by clinical evaluation to be affected with granular corneal dystrophy and carried the BIGH3 mutation W555R. However, both probands in families B and C, also diagnosed with granular corneal dystrophy, harboured the BIGH3 mutation R124H.</p><p><b>CONCLUSION</b>Molecular genetic analysis can improve accurate diagnosis of corneal dystrophy.</p>

Insulinllike growth factor 2 imprinting status and promoter usage in the placenta of macrosomia / 中华妇产科杂志

Objective To study the mechanism of marcosomia by investigating insulin-like growth factor 2(IGF_2)imprinting status,expression level and the promoter usage in the placenta of macrosomia. Methods We selected heterozygous cases for Apa Ⅰ polymorphism in exon 9 of IGF_2 gene and then analyzed its imprinting status in 168 placentas of macrosomia and normal pregnancies.IGF_2 transcription levels and promoter usages in macrosomic and normal placenta were evaluated by using semi-quantitative RT- PCR assay.Results Thirty specimens of macrosomic placenta and 30 of normal placenta were identified as heterozygous for IGF_2.All of the heterozygous specimens showed maintenance of imprinting.The expression of placental IGF_2 mRNA(2.2?1.2)was significantly higher in macrosomia than that of normal weight group (1.6?0.6,P 0.05).Conclusion It is possible that over expression of IGF_2 in placenta contributes to macrosomia while the promoter usage and imprinting status are not associated with macrosomia.

Genetic typing and antibiotic susceptibility testing of strains from pseudomonas aeruginosa keratitis patients / 眼科

Objective To study the characteristics of Genetic typing and the antibiotic susceptibility testing of strains from Pseudomonas aeruginosa keratitis patients.Design Experimental study,Participants 23 eyes of 23 patients of Pseudomonas aeruginosa keratitis.Methods The genomic was extracted and amplified with PCR.The PCR products were purified and sequenced.The results were registered in MIST web Antibiotic susceptibility testing were performed in theses strains,Main Outcome Measures Sequence types and antibiotic susceptibility.Results The isolates were resolved into 20 STs.Two lineages were identified.MIC test showed that strains were more susceptible to aminoglycosides,The activity of quinolones and cephalosporin were higher than that of aminoglycosides.Conclusion MIST can determine homology of the strains from Pseudomonas aeruginosa keratitis by clustering results. There is no finding about relationship between Genetic typing and drug resistance.

Study on the clinic and laboratory diagnosis of patients with trachoma / 眼科

Objective To investigate the clinical manifestation of trachoma,and to select the appropriate laboratory test for clini- cal diagnosis.Design Retrospective case series.Participants Retrospective analysis of medical records from 61 patients with trachoma from Jan 2003 to Aug 2006 in Bejing Tongren Eye Center.Methods Grades of trachoma diagnosis were according to the criteria de- signed by Chinese Ophthalmological Society (1979).The general state of health,case history,and the laboratory investigations of pa- tients were recorded.Laboratory tests included the conjunctiva scraping for inclusions,C.trachomatis immune antigen test and PCR test.Main Outcome Measures Manifestation of corneal and conjunctiva,the results of laboratory tests of C.trachomatis.Results Out of sixty-one patients including 28 males and 33 females,the average old was (29.05?19.99) years.88.5% cases were inⅠstage of tra- choma,8.2% were inⅡstage,and 3.3% were inⅢstage.The C.trachomatis inclusions were found in 7 (11.5%) scraping smears.42 (68.9%) cases were positive in C.trachomatis antigen test.46 (75.4%) cases were positive in PCR tests.The positive rates of antigen and PCR test were significantly higher than that of scraping (P=0.00).Conclusions A majority of clinical patients were inⅠstage of trachoma.The degree of their distress was minimal.It is necessary to apply C.trachomatis,antigen test or PCR test to improve the clin- ical diagnosis.

Clinical analysis about 5 cases of actinomycete keratitis / 眼科

Objective To analyze clinical diagnosis and management of 5 patients with actinomycete keratitis.Design Retro- spective case series.Participants 5 patients (5 eyes) with actinomycete keratitis.Methods The clinical features and microbiologic da- ta of 5 culture-proven cases of actinomycete keratitis recorded between October 2004 to March 2006 were analyzed.Main Outcome Measures clinical characteristics,isolations identification,drug susceptibility test and treatments.Results All patients were males and farmers.Of the 5 cases presented in this study,4 cases were followed by minor trauma as a predominant risk factor,and were pre- sented by a chronic progressive corneal ulcer with a wreath pattern of infiltrate.The diagnosis of all cases was based on laboratory in- vestigations,by which 4 cases of nocardia and one case of streptomyce were identified.A variable drug sensitivities were presented in nocardia isolates,which including TMP-SMZ,amicasin,gentamicin and fluorine-quinolones.Conclusions Nocardia keratitis is mainly followed by a minor trauma.It is identified predominantly by laboratory investigations.Tropical and systemically sensitive biotic are the initial choice,while debridement and amnionic transplantation could be an effective alternative.