AIM: To find out the gene structure and diversity of protate specific membrane antigen(PSMA) alternative spliced variants, and probe into the pathogenesis of prostate cancer.METHODS: 5'-RACE and 3'-RACE methods were used to amplify the 5' and 3'end of alternative spliced variant and then those viariants were sequenced for analyzing the gene stucture and diversity of PSMA alternative spliced variants of prostate cancer tissues.RESULTS: Four new alternative spliced variants of PSMA were discovered from prostate cancer tissues.Compared with reported PSMA alternative spliced variants,different insertions and deletions existed in different sites of those new variants.CONCLUSION: The discovery of the new variants confirms the diversity of PSMA spliced variants and provides the clues for seeking the target of diagnosis and therapy of prostate cancer.

Background Diabetic complication is associated with lipid peroxidation.Aldehyde dehydrogenases (ALDH) catalyze the irreversible oxidation of a variety of biological aldehydes,including lipid-derived aldehydes (LDAs),and thus protect organs and tissues from toxic LDAs.Understanding the activity of ALDH in different ocular tissues in diabetic subjects is very important for prevention and treatment of diabetic ocular complications.Objective This research aimed to investigate the activity and expression of ALDH in different ocular tissues in diabetic rats and to explore the mechanism of ALDH in diabetes-induced eye disease.Methods Twenty-eight healthy SPF male Sprague-Dawley(SD) rats weighted 170-180 g were randomly divided into the normal control group and diabetic group.The diabetic animal model was established by intraperitonial injection of 4％ streptozotocin at 65 mg/kg.Isometric citric acid buffer was injected in the rats of the normal control group.The rats were sacrificed in each group 2 and 4 months after the establishment of the diabetic models,and eyeballs were obtained for the preparation of corneal,lens and retinal homogenates.ALDH activity was detected using a multifunctional microplate reader SpectraMax M5,and ALDH content was measured by ELISA at the wavelength of 450 nm with the SpectraMax M5 ELISA reader.Results The blood glucose level in diabetic rats was significantly elevated at various time points compared with the normal control group(P=0.000),and body weights were evidently lower in the diabetic group than in the normal group (P =0.000).The activities of ALDH (A340) in corneal,lens and retinal tissues in the diabetic group were increased in comparison with the normal control group (F =396.601,P=0.000),and showed an enhancement with the lapsing of time (F =53.139,P =0.000).In addition,the highest level of ALDH was found in the cornea and the lowest level in the lens(F =6973.000,P=0.000).The expression level of ALDH in the corneal,lens and retinal homogenates was significantly higher in the diabetic group compared with the normal control group (F=312.985,P =0.000) and showed a considerable increase over the course (F =19.203,P=0.000).The highest expression level was seen in the cornea and the lowest was in the lens,with a significant difference among these three kinds of tissues (F =3243.000,P =0.000).Conclusions ALDH can protect ocular tissue from the damage of lipid peroxidation.Thess results suggest that ALDH plays a role in preventing diabetes-related ocular complications.

Background Subretinal transplantation of retinal pigment epithelium (RPE) cells for the treatment of age-related macular degeneration (AMD) have accelerated the drive to develop xeno-free cultivation system that support the rapid differentiation of human embryonic stem cells (hESCs) into ES-RPE cells.Objective This study was to report a modified xeno-free culture system and method for accelerating derivation of hESCs to differentiate into RPE cells.Methods This study was approved by Ethic Committee of Zhongshan Ophthalmic Center.HESC H1 line was cloned and cuhured in Vitronectin XFTM-coated 6-well dish with xenogenetic-free medium.Cells were cultured in 50 ng/ml noggin,10 ng/ml DKK-1 and 10 ng/ml insulin like growth factor-1 (IGF-1) medium for 2 days,and then the concentration of noggin was decreased to 10 ng/ml and 5 ng/ml basic fibroblast growth factor (bFGF) and cultured for the following 2 days.Sequentially,noggin and bFGF were removed and cultured for 2 days.Finally,1 μmol/L CHIR99021 was added in medium for 6 days.Morphological changes in the progress of ESCs differentiation into RPE were observed by Living Cell Imaging System.The expression of Mitf and RPE65,RPE cellsspecific markers,in the cells were detected by immunofluorescence technique,and the relative expression levels of RPE cells-specific marker mRNA were assayed using real time fluorescent quantitation PCR.Results Polygonalshape monolayer cells which contained pigments were initially observed at day 14 after cultured with the cobblestonelike arrangement.Mitf and RPE65 were strongly expressed in the hES-derived RPE cells 35 days after induced,showing red fluorescence,and the cells presented hexagonal shape at cultured day 60 with numerous pigment granules in cytoplasm.Compared with before differentiation,the expression levels of Mitf mRNA in hES-RPE cells increased by (3.43±2.77) folds and (8.91 ± 2.83) folds,and the expression levels of RPE65 mRNA increased by (14.60 ± 3.94) folds and (87.16 ±9.32) folds at day 7 and day 14 after differentiation,respectively (all at P＜0.05).Conclusions A defined xeno-free culture system is successfully established by adding niacinamide,DKK-l,noggin,IGF-1 and CHIR99021 in xeno-free medium,and this system can accelerate the derivation and differentiation of hESCs into RPE-like cells.

Objective Hypertension is a leading modifiable risk factor for cardiovascular disease .However , a lot of hyper-tension patients hold inactive attitudes to hypertension treatment .The purpose of this study was to investigate the relationship between previous treatment of hypertension and stroke severity in acute ischemic stroke . Methods We retrospectively analyzed the clinical data of 653 in-hospital ischemic stroke patients with hypertension between January 2011 and December 2014 .According to the National Institutes of Health Stroke Scale (NIHSS) at admission, the stroke patients were divided into a mild group (NIHSS≤3) and a severe group (NIHSS >3) and, based on their history of hypertension treatment , allocated to a regular treatment, an irregular treatment, a non-treatment , and an unawareness group .We studied the relationship of previous hypertension treatment with stroke severity by Spearman correlation analysis and identified the potential factors associated with stroke severity by multivariate logistic regression anal-ysis. Results Previous treatment of hypertension was positively correlated with stroke severity (r=0.146, P=0.000 2).Compared with the patients of the regular treatment group , those in the irregular treatment group (OR: 2.21; 95%CI:1.39 -3.52; P =0.001), non-treatment group ( OR: 2.18; 95%CI: 1.41 -3.36; P =0.0004) and unawareness group (OR:1.80;95%CI:1.12-2.88; P=0.015) tended to have more severe stroke. Conclusion Previous treatment of hypertension is closely related to the severity of ischemic stroke .

Identiifcation of tissues/body lfuids in forensic science is important for criminal cases investigation such as crime scene reconstruction, conclude the character of crime. Recently, many researches of Epigenetic shows that tissue speciifc differentially methylated regions(tDMRs) have the ability to as a biomarker for identiifcation of tissues/body lfuids. In this paper, we reviewed the study progress and summarized the probability, advantage and disadvantage as well as application value and the development direction of the application of DNA methylation in the aspect of identifying the tissues/ body lfuids source, aiming at providing a reference for the related research and application.

The study compared the traditional medical microbiology experimental teaching with a new experimental teaching pattern,with the students majoring in anaesthesia as the research object. The new pattern mainly deals with the cultivation of the students' creativity by reforming and exploring the plan,the content,the method of experimental teaching and the ways of checking the students' work, adding general and designing experiment,and working out the PPT of the experiments.The result shows the new experimental teaching pattern contributes to the cultivation of the students'abilities of performing experiment, the ways of thinking, creativity and comprehensive analysis. It's better than the traditional experimental teaching pattern.A new medical microbiology experimental teaching systerm which is suit to the students majoring in anaesthesia has been established.

Objective Meningovascular syphilis is the intima inflammation of blood vessels caused by the syphilitic infec-tion, which is associated with the occurrence of ischemic stroke.The study analyzed the clinical, imaging features and prognosis for meningovascular syphilis so as to improve its diagnosis and treatment. Methods 14 patients diagnosed with meningovascular syphilis were collected prospectively from December 2007 to March 2015 in the neurological department of Jinling Hospital.The patients were followed for a period of 21.5(range 10.2~37.9)months,and the prognosis were evaluated. Results Patients with meningovascular syphilis presented with dizziness, hemiplegia, hemidysesthesia and cognitive decline.Lesions showed multiple, scattered on MR ima-ging, intracranial vascular stenosis was seen in the CTA/MRA, and the laboratory examination had characteristic changes.With a large dose and sufficient courses of penicillin treatment, meningovascular syphilis may hopefully get predominant effects.78.57%patients got good prognosis(modified Rankin Scale ,mRS≤2)at 3 months and 85.71% patients got goodlong-term prognosis(mRS≤2). Conclusion Meningovascular syphilis was usually presented as acute onset, lacks the specific clinical and neuroimaging manifesta-tions.Most patients has favourable prognosis after treatment of syphilis with full course of penicillin.

Objective To analyze the functional changes and the clinical significance of B cell specific mono-clonal murine leukemia virus integration site -1 (Bmi -1 )and Th1 /Th2 cells in children with newly diagnosed im-mune thrombocytopenia(ITP)by testing the mRNA expressions of Bmi -1,helper T cell -related cytokine interferon (IFN)-γand interleukin(IL)-4 in children with newly diagnosed ITP.Methods Thirty -six cases of patients with newly diagnosed ITP in the experimental group came from the inpatient and outpatient children admitted to the Depart-ment of Pediatrics of the First Affiliated Hospital of Xinxiang Medical University from April to December 201 3.In the control group,26 cases of children requiring selective operation were admitted to the Department of Pediatric Surgery during the same period.The mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes were detected by means of the reverse transcription -polymerase chain reaction(RT -PCR)method,and were analyzed and compared by t test and linear correlation analysis.Results (1 )The mRNA expressions of Bmi -1,IFN -γand IL -4 in peripheral blood lymphocytes in the experimental group were 2.63 ±0.54,3.84 ±0.43 and 1 .44 ±0.39,respec-tively;while the mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes in the control group were 3.91 ±0.92,2.88 ±0.57 and 1 .87 ±0.34,respectively.The levels of IFN -γof the experimental group were significantly higher than those of the control group (P <0.001 )and the levels of Bmi -1 and IL -4 in the experi-mental group were significantly lower than those in the control group (all P <0.001 ).(2)The mRNA expressions be-tween IFN -γand IL -4 in the peripheral blood lymphocytes in the experimental group were in negative correlation (r =-0.667,P <0.001 ).The mRNA expressions between IL -4 and Bmi -1 in the same group were in a positive correlation (r =0.776,P <0.001 ).There were no correlation in the mRNA expressions between IFN -γand Bmi -1 (r =-0.206,P >0.05).Conclusions Bmi -1 may be involved in the pathogenesis of ITP by regulating Th cell, and Th cell dysfunction may occur in the children with ITP,and the disproportion between Th1 and Th2 may be due to the advantages of Th1 .

Objective:To investigate the effects of oxymatrine(OM)on the apoptosis of human hepatoma HepG2 cells and its possible mechanisms.Methods:HepG2 cells were treated with different concentrations of OM.The proliferation inhibition was measured by MTT assay and the apoptosis of HepG2 cells were examined by Hochest staining method.Flow cytometry was used to analyze the cell cycle distribution and apoptosis rate.The expression of caspase-3,Bcl-2,Bcl-x_L and Bax proteins was assayed by Western blotting assay.Results:OM inhibited HepG2 cells growth in a time-and dose-dependent manner.After treatment with OM for 24 hours,some cells appeared typical apoptotic characteristics and the apoptosis rate was increased. Treatment with OM also increased caspase-3 activity and Bax expression in HepG2 cells,and decreased the expression levels of Bcl-2 and Bcl-x_L.Conclusion:OM can induce HepG2 cell apoptosis,which may be related to the down-regulation of PI3K/Akt signal pathway,suppression of Bcl-2 and Bcl-x_L activity,and activation of caspase-3.

Objective To explore the effects of forepaw sensorimotor deprivation in early life on hippocampus-dependent spatial reference learning and memory in rats. Methods Newborn SD rats were randomly assigned to experiment group (deprivation of forepaw sensorimotor function, n=53) and control group(n=55). Rats of postnatal day 13 (PN13) in experiment group were seleeted, and models of forepaw sensorimotor deprivation were established by microsurgical technique. Open field tests and Morris water maze tests were performed during the time periods of PN25(PN21-31), PN35 (PN31-39), PN45(PN41-50) and PN60(PN56-64) to evaluate the locomotor activity and spatial reference learning and memory, respectively. Results In open field tests, there was no significant difference in parameters of locomotor activity and exploratory behavior between the two groups (P>0.05). In Morris water maze tests, eontrol group performed significantly better than experiment group during training sessions and probe tests on PN25 and PN35 (P<0.05). While on PN45, although there was no significant difference between the two groups during training sessions, control group performed significantly better than experiment group during probe tests (P<0.05). Conclusion The deprivation of forepaw sensorimotor in early life has no signifieant effect on the locomotor activity and exploratory behavior of rats, but can impair the spatial reference learning and memory.