At present,the fashions,methods and means of medical genetics teaching have fallen behind the development of the subject.Under the patterns of net-teaching,the construction of net-teaching database is an important tache to develop teaching favourably,improve the quality and efficiency of teaching fleetly.This paper introduces the process of constructing a net-teaching database for medical genetics,the applications and characters of the database in teaching.

OBJECTIVETo study the roles of stromal interaction molecule 2 (STIM2) and transient receptor potential canonical 3 (TRPC3) in extracellular Ca(2+)-sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO) in human umbilical vein endothelial cells (HUVEC).

METHODS(1) The interaction of STIM2 and TRPC3 was determined using the immunofluorescence technique. (2) The expressions of STIM2 and TRPC3 genes were silenced in HUVEC by transfection constructed STIM2 and TRPC3 RNA interference plasmids. The interference efficiency of STIM2, TRPC3 protein and mRNA levels were determined by Western blot and real time RT-PCR, respectively. (3) The second to fifth passage of HUVEC were divided into: STIM2-002 short hairpin RNA (STIM2-002 shRNA ) + spermine + Ca2+ group and TRPC3-004 short hairpin RNA (TRPC3-004 shRNA ) + spermine + Ca2+ group; control group (spermine + Ca2+ group) and vehicle+ spermine + Ca2+ group. The four groups of cells were incubated with CaR agonist spermine, the intracellular Ca2+ concentration ([Ca2+]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, and the production of NO was determined by DAF-FM (NO fluorescent probe) of each group in HUVEC.

RESULTS(1) Immunofluorescence technique results showed that STIM2 and TRPC3 proteinswere present in the cytoplasm of HUVEC. (2) The results of transfection constructed STIM2 and TRPC3 RNA interference plasmids demonstrated that shRNA targeted to the STIM2 and TRPC3 genes decreased STIM2 and TRPC3 mRNA levels by 88.2% and 74.0%, respectively (P < 0.05), simultaneously, the STIM2 and TRPC3 protein levels were decreased by 79.9% and 71.8%, respectively (P < 0.05). (3) Compared with spermine + Ca2+ group, the [Ca2+]i and the net NO fluorescence intensity of spermine + Ca(2+) + ShSTIM2-002 group, spermine + Ca(2+) + ShTRPC3-004 group and spermine + Ca2+ Vehicle group were not changed (P > 0.05).

CONCLUSIONSTIM2 and TRPC3 do not participate in CaR-mediated Ca2+ influx and NO production individually.

Calcium ; metabolism ; Cell Adhesion Molecules ; physiology ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; physiology ; Humans ; Nitric Oxide ; metabolism ; Stromal Interaction Molecule 2 ; TRPC Cation Channels ; physiology

Objective To investigate the role of CD4~+CD25~+ Treg cells on the pathogenesis of ankylos- ing spondylitis(AS), and to study the machanism of tumor necrosis factor(TNF)-?blocker on the treatment of AS by detecting the number of CD4~+D25~+ Treg cells before and after the treatment. Methods The diagno- sis of 10 AS patients was made based on the 1984 modified New York criteria. The patients received subcuta- neou injection of recombinant human tumor necrosis factor receptor-Fc fusion protein(rhTNFR-Fc)(etaner- cept)50 mg weekly for 8 weeks and 10 heathy subjects were enrolled for control. The mononuclear cells were isolated from peripheral blood in beth patients and controls. The number of CD4~+CD25~+ T cells and CD4~+ CD25~(high)T cells and the expression of CTLA-4. were detected by flow cytometry. Results The proportion of CD4~+CD25~+ T cells(24?19)% in total CD4~+ T lymphocytes of peripheral blood and CD4~+CD25~(high)T/CD4~+ T (6?6)% from AS patients before treated with rhTNFR-Fc was higher than that in healthy volunteers and AS patients after treatment(P

Background Neovascular glaucoma (NVG) is a serious ocular disease which may cause blindness.The primary pathogenesis of NVG is ischemic retinopathy derived by central retinal vein occlusion (CRVO) and diabetic retinopathy (DR).Clinical characteristics of NVG are variable based on the difference of primary diseases,such as CRVO and DR.However,there is a few studies regarding the diffcrcnces of NVG initiated by CRVO and DR.Objective This study was to compare the clinical characteristics in NVG patients secondary to CRVO and DR.Methods A series case observational study was carried out in Hiserve Hospital of Qingdao University from January 2009 to June 2012.Twenty-nine eyes of 27 patients with NVG caused by CRVO (10 eyes of 10 patients) and DR (19 eyes of 17 patients) were included.The history of underlying diseases,course of NVG,intraocular pressure(IOP),fundus findings and complications after treatment were analyzed and compared between the CRVO-derived NVG and DR-derived NVG.All patients underwent panretinal photocoagulation,improving microcirculation therapy,anti-glaucoma (drug or surgery) and causative disease treatment,and some of them received vitrectomy or/and cataract surgery.Two eyes from each group received intravitreal injection of ranibizumab.The follow-up time in both groups was (14.00±10.13) months and (17.89±12.52) months,respectively.Results The median time of underlying disease was 3.3 months (2 weeks to 6 months) in the CRVO patients and 11.1 months (4 to 36 mouths) in the DR patients,with a significant difference between them (Z =-2.40,P＜0.05).CRVO-derived NVG progress was much faster than that of DR-derived NVG.The number of the eyes with visual acuity improvement after treatment was 2 in the CRVO-derived NVG and 15 in the DR-derived NVG;while the number of the eyes with unchanged or worse visual acuity was 8 and 4 in the CRVO-derived NVG eyes and the DR-derived NVG eyes (x2 =9.38,P＜0.01).The difference of IOP in pre-and post-treatment was (37.00±9.91)mmHg in the CRVOderived NVG eyes and (8.92±12.05)mmHg in the DR-derived NVG eyes,showing a significant difference between them (t =6.30,P＜0.01).In the CRVO-derived NVG eyes,optic disc edema,retinal hemorrhage,and vein dilatation were seen in 6 eyes,and mild optic disc edema and retinal hemorrhage were observed in 4 eyes.After treatment,fundus could not be seen in 4 eyes,in other 2 eyes optic disc and retinal laser spots were unclearly observed.In addition,pale optic disc and retinal vessel occlusion appeared in 2 eyes,and silver wire-like arteries exhibited in 2 eyes.In pre-treated DR-derived NVG eyes,fundus could not be seen in 8 eyes and Ⅲ-Ⅳv stages of DR findings appeared in 11 eyes.After treatment,retinopathy was stabilized in 16 eyes of 15 cases.Advanced retinopathy(V-Ⅵ stages of DR findings) was revealed in 3 eyes of 3 cases.The incidence of the complication after treatment was 100.0％ in the CRVO-derived NVG eyes and 21.1％ in the DR-derived NVG eyes (x2=5.18,P＜0.05).Conclusions The clinical characteristics of NVG secondary to CRVO and DR are variable,an appropriate treatment option should be selected according to different features of NVG.

OBJECTIVETo observe the differences of therapeutic effects on common cold of wind-cold type at early stage treated by different cupping duration.

METHODSSixty cases of common cold of wind-cold type were randomly divided into an observation group and a control group by fifty-fifty. Flash cupping was applied at Dazhui (GV 14), Fengmen (BL 12), Feishu (BL 13), Xinshu (BL 15) and Geshu (BL 16), etc., and the cupping was retained for 25-30 min in the observation group and 15 min in the control group. The therapeutic effects were compared after twice treatments.

RESULTSThe total effect rate was 93.3% (28/30) in observation group, superior to that 66.7% (20/30) in control group (P < 0.01).

CONCLUSIONThe therapeutic effect of common cold of wind-cold type at early stage treated with cupping for 25-30 min is obvious, superior to that for 15 min.

Acupuncture Points ; Adolescent ; Adult ; Common Cold ; therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Treatment Outcome ; Young Adult

The flavonoids were investigated from the whole plants of Lagopsis supina. The compounds were isolated and purified by various column chromatography, and their structures were identified by physiochemical properties and spectroscopic data. Two flavones were isolated from the CH2Cl2 layer of Lagopsis supina extract and identified as genkwanin (1) and 5-hydroxy-7,4'-dimethoxyflavone (2), respectively. Ten flavonoid glycosides were isolated from the water layer of Lagopsis supina and elucidated as kaempferol-3-O-6" (3-hydroxy-3-methylglutaryl) -β-D-glucoside (3), quercetin-3-O-6"-(3-hydroxy-3-methylglutaryl) -β-D-glucoside (4), quercetin-3-O-β-D-glucoside(5), kaempferol-3-Of3-D-glucoside ( 6), isorhamnetin-3-O-/-D-glycopyranoside (7), apigenin-7-O-6-D-glucoside (8), luteolin-7-O-β-D-glucoside (9), chrysoeriol-7-O-β-D-glucoside (10), rutin (11 ), and kaempferol-3-β-(6"-p-coumaroyl) -β-D-glucoside (tiliroside, 12). Compounds 3 and 4 were firstly isolated from Lamiaceae family, and compounds 1-12 were isolated from the plants of Lagopsis genus for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

AIMTo investigate the relationship between low androgen level and ultrastructure of vascular endothelium.

METHODSForty-eight male Sprague-Dawley rats were randomly divided into four groups: group A, normal rats with sham castration; group B, castrated rats; group C, castrated rats given testosterone (T) undecanoate; and group D, intact rats treated with 5alpha-reductase inhibitor. After 10 weeks of treatment or castration, rats in different groups were killed and serum T, free T (FT) and dihydrotestosterone (DHT) were measured. The aortic endothelia were scanned under electron microcopy and the Vascular Endothelium Structure Score (VESS) was computed.

RESULTSSerum T and FT concentrations of rats in group B were significantly lower than those of the other three groups (P < 0.01); DHT concentrations of group D rats were significantly decreased (P < 0.01) when compared with those of groups A and C. Rats in groups B and D rats (with low androgen levels) had obvious damage to their endothelial surfaces, which appeared crimpled, rough, adhesive and ruptured, and had high destruction of VESS.

CONCLUSIONThese results suggest that low concentrations of T and DHT are associated with ultrastructural damage of the aortic endothelia in male rats.

5-alpha Reductase Inhibitors ; Animals ; Aorta ; drug effects ; ultrastructure ; Dihydrotestosterone ; blood ; Endothelium, Vascular ; drug effects ; ultrastructure ; Enzyme Inhibitors ; pharmacology ; Male ; Orchiectomy ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood ; pharmacology

OBJECTIVETo explore the relationship between the polymorphism of estrogen-biosynthesis genes (CYP17, CYP19, HSD17beta1) and risk of breast cancer.

METHODSA matched case-control study was designed. From May 2007 to July 2008, 200 pairs of subjects with and without breast cancer were enrolled, who were matched by age and menstruation status. Demographical characteristics, dietary factors and reproductive factors were investigated by questionnaire. CYP17 locus 1931 (T-->C), CYP19 codon 264 (Arg-->Cys) and HSD17beta1 locus 1954 (A-->G) were identified by AS-PCR (allele-specific PCR). The gene-gene interaction were analyzed with the MDR model (multifactor dimensionality reduction). Based on the results of MDR model, an unconditional logistic regression model was simulated to estimate the ORs of interaction factors and other risk factors.

RESULTSThe main effect of CYP17, CYP19 and HSD17beta1 susceptible genotypes were not correlated to breast cancer (OR approximately 1, P > 0.05). The positive interaction effect between CYP17 (T 1931C) and HSD17beta1 (A1954G) was discovered by MDR model with a statistically significant difference (Sign test, P = 0.05). The model's testing balance accuracy was 56.00%, and crossing validation consistency was 10/10. Multivariable unconditional logistic regression showed that after adjusting BMI, intake of estrogen, age of first birth, number of abortion and period of breast feeding, the interaction item of CYP17 (T1931C) and HSD17beta1 (A1954G) was strongly and positively correlated to breast cancer (OR = 2.52, 95%CI = 1.54 to 4.11).

CONCLUSIONThe estrogen-biosynthesis genes CYP17 (T1931C) and HSD17beta1 (A1954G) polymorphism may jointly increase the risk of breast cancer.

Aromatase ; genetics ; Breast Neoplasms ; genetics ; pathology ; Carcinoma, Ductal, Breast ; genetics ; pathology ; Case-Control Studies ; Estradiol Dehydrogenases ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Logistic Models ; Polymorphism, Single Nucleotide ; Risk Factors ; Steroid 17-alpha-Hydroxylase ; genetics

This study was aimed to investigate the expression of RHBDD1 gene in patients with chronic myeloid leukemia (CML) and explore its clinical significance. The relative expression levels of RHBDD1 in bone marrow mononuclear cells of healthy controls and CML patients were detected by using real time PCR. The results showed that the expression level of RHBDD1 in CML patients was significantly higher than that in healthy controls. The expression level of RHBDD1 in CML patients with negative BCR/ABL p210 was remarkably higher than that in patients with positive BCR/ABL p210. In patients ≥ 50 years old RHBDD1 expression was lower than the patients < 50 years old. There were no significant relation of RHBDD1 expression with sex of patients. It is concluded that RHBDD1 gene may be involved in the pathogenesis and progression of CML, particularly reflects in the pathogenesis of the patients with negative BCR/ABL p210.
Bone Marrow Cells ; metabolism ; pathology ; Case-Control Studies ; Female ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Serine Endopeptidases ; genetics ; metabolism

Larp4b is a member of the LARP family, which can interact with RNA and generally stimulate the translation of mRNA. Abnormal expression of Larp4b can be found in leukemia patients in our previous study. This study was purposed to detect the relative expression of Larp4b mRNA in different subpopulations of mouse hematopoietic cells, to construct lentivirus vector containing shLarp4b targeting mouse gene Larp4b and to explore its effects on mouse Lin(-) cells infected with shLarp4b by lentivirus. SF-LV-shLarP4b-EGFP and control vectors were constructed and two-plasmid lentivirus packing system was used to transfect 293T cells. After 48 h and 72 h, lentivirus SF-LV-shLarp4b-EGFP was harvested and was used to infect Lin(-) cells. After 48 h, EGFP(+) cells was sorted by flow cytometry (FCM). Meanwhile, semi-quantitative real time-PCR, AnnexinV-PE/7-AAD staining, PI staining and colony forming cell assay (CFC) were performed to determine the expression of Larp4b and its effect on the proliferation of hematopoietic progenitor cells. The results showed that Larp4b was highly expressed in myeloid cells. SF-LV-shLarp4b-EGFP was successfully constructed according to the restriction endonuclease digestion assay. RT-PCR confirmed that Larp4b was efficiently knockdown in mouse Lin(-) cells. The low expression of Larp4b did not affect the colony forming number, the apoptosis and cell cycle of Lin(-) cells. It is concluded that knockdown of Larp4b in mouse Lin(-) cells do not contribute to the colony forming ability and the growth of Lin(-) cells in vitro. This useful knockdown system will be used to study in vivo Larp4b in future.
Animals ; Autoantigens ; metabolism ; Cells, Cultured ; Flow Cytometry ; Gene Knockdown Techniques ; Genetic Vectors ; Hematopoietic Stem Cells ; cytology ; Humans ; Lentivirus ; genetics ; Mice ; Plasmids ; Ribonucleoproteins ; metabolism ; Transfection