Objective To confirm the transmission of Toxoplasma gondii by semen and to investigate the impact of vaginal status on the transmission of T. gondii in female rabbits. Methods Sixteen male rabbits were infected with T. gondii by intraperitoneal injection each with 1 ×105 RH tachyzoites. Eight rabbits died in 8-14 d after infection.Artificial vagina was used to collect semen from male rabbits weekly before and after infection for 8 weeks. If more than 2 portions of semen from 8 survived male rabbits were collected after infection, the collected semen was mixed weekly for later use. Twenty-seven female rabbits were divided into 4 groups: group 1 with normal vagina (7 rabbits), group 2with wounded vagina (7), group 3 with trichomonas vaginitis (7) and group 4 with colpomycosis infection (6). Tachyzoites were found in mixed semen digested by trypsinase, and were used for endovaginal artificial insemination to female rabbits by uterine cavity tube once a week for 8 consecutive weeks. 2-3 d after every insemination, 2 ml blood was collected from helix vein of each rabbit, and stored at -40 ℃ for use. Anti-T. gondii antibody was examined by ELISA and the B1 gene of T. gondii was detected by PCR. Results Anti-T. gondii antibody was detected in some rabbits (2, 3, 1, and 1 rabbits from each of the groups respectively) on the 16th day after the first insemination. The positive rate of ELISA was 25.9%. The amplification of B1 gene (200 bp) by PCR appeared positive from the blood samples on the 3rd day after the first insemination and the last positive one was proved on the 51th day after the first insemination.Number of positive samples was 2, 1, 3 and 1 in the 4 groups respectively, with an overall PCR positive rate of 18.5%.Only 3 of the 27 rabbits were positive by both ELISA and PCR. Conclusions T. gondii can be transmitted by semen and the health status of vagina shows no impact on it.

Objective To investigate the association between single nucleotide polymorphism (SNP) of macrophage migration inhibitory factor (MIF) gene-rs1007888 and the pathogenesis of gestational diabetes mellitus (GDM).Methods A total of 120 GDM pregnant women (GDM group) and 165 healthy pregnant women (control group) from Affiliated Hospital of Medical College,Qingdao University were recruited from June 2011 to July 2012.Their age,gestational week,height and weight were recorded.The levels of fasting blood glucose (FBG) and fasting insulin (FIN) were determined.Body mass index (BMI),the hemeostasis model assessment-insulin resistance (HOMA-IR) and hemeostasis model assessment-β cell function (HOMA-β) were calculated.DNA was extracted from fasting blood samples.SNP of MIFrs1007888G/A was determined by DNA sequencing.The FBG,FIN,HOMA-IR and HOMA-β were compared between GDM group and the control group.They were also compared among pregnancies withdifferent genotypes.Results (1) GDM group had higher FBG,FIN and HOMA-IR levels,but lower HOMA-β than the control group (all P ＜ 0.05).(2) MIF-rs1007888 SNP genotype frequencies of GG,GA and AA were 37.5％,45.8％ and 16.7％,and the allelic frequencies of G and A were 60.4％,39.6％ in GDM group; However,in the control group,the frequencies of GG,GA and AA were 26.1％,54.5％ and 19.4％,and the allelic frequencies of G and A were 53.3％,46.7％,respectively.The distributions of MIF genotypes in GDM patients were significantly different from the healthy subjects (P ＜ 0.05).No significant difference of MIF-rs1007888 allele distributions was observed between GDM group and the control group (P ＞0.05).(3) The FBG,FIN and HOMA-IR in pregnant women with GG genotype were statistically higher than those with GA or AA genotypes,while HOMA-β was lower in women with GG genotype (all P ＜0.05).Conclusions The SNP of MIF rs-1007888 was related to the insulin resistance and pancreatic β cell function of pregnant women.GG genotype of MIF-rsl007888 might be a genetic susceptible factor in the pathogenesis of GDM.

Objective To study the mutation of STK11 gene in a Chinese family and a sporadic patient with Peutz-Jeghers syndrome (PJS),and to provide a basis for genetic diagnosis and counseling.Methods One sporadic patient and two patients from a family with PJS were collected,all of whom had typical mucosal pigmentation and gastrointestinal polyposis.Blood samples were obtained from the two patients and six unaffected relatives in this family,the sporadic patient,and 100 healthy controls.DNA was extracted,and PCR was performed to amplify nine exons and their adjacent introns in the STK11 gene followed by direct sequencing.The sequencing results were aligned to the published sequence of STK11 gene from Genbank.Results No mutation was found in the STK11 gene of any of the patients,unaffected relatives,or healthy controls.Conclusions Genetic heterogeneity exists in Peutz-Jeghers syndrome,hinting that there may be other causative genes or sites for this entity.

OBJECTIVE To introduce a new type of tracheal cannula which has the top hole and can seal the outlet for patient with severe brain injury after tracheotomy. METHODS Patients with severe brain injury after tracheotomy were divided into test group using the new type of tracheal cannula(25 cases) and control group using the cuffed Portex tube(25 cases).The incidence,the onset time and the time for control of pulmonary infection were compared between the two groups. RESULTS In the test group,the incidence of pulmonary infection was 52%(13 cases),the onset time was(3?1.3) days and the time for control was(5?1.7) days;in the control group,the above indices were 84%(21 cases),(3?1.5) days and(12?2.2) days separately. CONCLUSIONS The new type of tracheal cannula which has the top hole and can seal the outlrt is better than the cuffed Portex tube in the prevention and treatment of pulmonary infection after tracheotomy.

Objective To establish a RP- HPLC method for quantitative determination of puerarin in Laige Granules. Methods The separation was performed on Econosphere C18 column (4.6 mm? 250 mm,5 ? m) with mobile phase of methanol and 1 % acetic acid solution (25 ∶ 75) at a flow rate of 0.8 mL/min. The UV detection wavelength was 250 nm and the column temperature was 40 ℃ . Results The linear range of puerarin was 0.2～ 1.0 ? g, r = 0.999 7. The mean recovery was 100.56 % (RSD=1.21 % , n = 5 ). Conclusion This method is simple, accurate, sensitive and with good reproducibility .

Objective To investigate the genotypic and allele frequency differences of melatonin receptor 1B (MTNR1B)-rs4753426 between gestational diabetes mellitus (GDM) pregnancies and normal pregnancies , and to explore the association between single nucleotide polymorphism ( SNP ) of rs4753426 and gestational diabetes mellitus.Methods Totally 93 GDM pregnancies and 165 normal pregnancies were recruited from the Affiliated Hospital of Qingdao University.The age, gestational weeks, height, early pregnant weight , and the levels of fasting plasma glucose ( FPG) , fasting insulin ( FIN) were determined in every participants.By using PCR and DNA sequencing , we detected the distribution of the rs 4753426 genotypes and alleles in all individuals.The homeostasis model assessment-insulin resistance ( HOMA-IR) and the homeostasis model assessment-βcell function ( HOMA-β) were calculated.The allele and genotype frequencies and the FPG , FIN, body mass index ( BMI) , HOMA-IR, HOMA-βlevels between GDM group and control group were compared.Results (1) The genotype frequencies in the GDM group and the control group of rs4753426-CC, CT, TT were 72.0% (67/93), 21.5% (20/93), 6.5% (6/93), and 53.9%(89/165), 40.0% (66/165), 6.1% (10/165) respectively.The allele frequencies in the GDM group and the control group of T and C were 17.2% ( 32/186 ) , 82.8% ( 154/186 ) and 26.1% ( 86/330 ) , 73.9% ( 244/330 ) respectively.There were statistical differences in genotype frequencies and allele frequencies between two groups ( all P<0.05 ).( 2 ) The levels of FPG , FIN and HOMA-IR in the GDM group were obviously higher than those in the control group (P<0.05).The level of HOMA-βwas lower in the GDM group than that of the control group (P<0.05).(3)The FPG of CC and CT genotypes was higher than that of TT genotype in the GDM group (P<0.05), while the level of HOMA-βwas lower than that of TT genotype (P<0.05).Conclusions The MTNR1B-rs4753426 SNP is associated with the pathogenesis of GDM, and rs4753426 is the predisposing locus of GDM.The C-allele is the susceptibility allele of GDM.

Objective To investigate whether polymorphism of 102 T/C in 5-HTR2A (serotonin receptor 2A) are associated with Tourette syndrome (TS) in Chinese Han population or none.Methods A total of 101 TS patients and their parents were recruited for the study.The genetic contributions of the 5-HTR-2A 102 T/C polymorphism in 5HTR2A were evaluated using polymerase chain reaction and restriction enzyme digestion (PCRRFLP) and haplotype relative risk (HRR) and transmission disequilibrium test (TDT) statistics.Results The results revealed no significant associations between the 5-HTR-2A 102 T/C polymorphism and TS (HTR-2A 102T/C,TDT =0.353,df=1,P =0.621 ;HRR =1.127,x2 =0.358,P =0.550,95％ CI:0.762-1.666).Conclusion The data suggest that the HTR-2A 102 T/C polymorphism may not be associated with susceptibility to TS in the Chinese Han population.However,these results need to be replicated using larger datasets collected from different populations.

Objective To study the increased radiosensitivity of tumor cells by triterpene acids of loquat leaf (TAL) and mechanism.Methods C6 and MG63 cells were pretreated by TAL,and then were exposed to X-rays at 1,2,3,5,7 Gy.Clonogenic assay was used to evaluate those tumor cells survival fraction (SF) to calculate the sensitization enhancement ratios (SER) of TAL.The cellular micronuclei ratios of tumor cells were analyzed by micronuclei assay.Additionally,the changes of Rad51 and XRCC4 levels were observed by RT-PCR and Western blot.Results D0 values were decreased to 1.31 and 2.85 Gy in C6 and MG63 cells by TAL pretreatment respectively.The SER value of the effect of TAL on C6 and MG63 cells was 1.73 and 2.04,respectively.There was a statistical difference in the cellular micronuclei ratios between tumor cells with TAL above 2 Gy and those without TAL (C6:t =-8.372--2.476,P ＜0.05 ; MG63:t =-4.03--2.557,P ＜ 0.05).TAL attenuated the expression of XRCC4 at transcriptional and translational level,but not for Rad51,a key gene in homologous recombination repair (HRR).Conclusions TAL pretreatment could increase the lethal effect of X-rays on tumor cells in vitro.The mechanism might be involved in the inhibition of non-homologous end joining (NHEJ).

Objective To explore gene polymorphism of G/A genotype of Fok Ⅰ rs2228570 (G/A) of vitamin D receptor (VDR) gene in Han male population of Chinese coastal area,and thus to investigate the relationship between the gene polymorphism of VDR and gout.Methods Altogether 504 gout patients and 523 healthy controls were enrolled.The possible association between the polymorphism of VDR rs2228570 and gout in Chinese coastal area was investigated and genotype frequencies and allelic frequencies were calculated by realtime PCR with Taqman(R)probe method.Hardy-Weinberg was used to verify the representativeness of the sample.Comparison between the groups were performed withx2 test and t-test.Results The frequencies of GG,AG,and AA genotypes were 32.1％,50.0％,and 17.9％,respectively among gout patients,while they were 27.9％,50.5％,and 21.6％ respectively among the controls.There was no statistically significant difference in VDR rs2228570 genotype frequencies between gout patients and controls(x2 =3.366,P＞0.05).The allele frequencies of G and A in gout cases were different from those in the controls(57.1％,42.9％;53.2％,46.8％;x2 =3.300,P＞0.05).Conclusions Results of the present study suggest that the G/A genotype of VDR Fok Ⅰ rs2228570 of the VDR gene is not associated with gout in male population of Chinese coastal area.

Eighty-two newly-diagnosed type 2 diabetic patients with poor glycemic control were treated by mitiglinide calcium for 16 weeks.Plasma fibroblast growth factor-21 ( FGF-21 ) level were evaluated.The relationship of plasma FGF-21 levels with body mass index,body fat,waist-to-hip ratio,lipid,blood glucose,HbA1c,and free fatty-acid were analyzed.Plasma FGF-21 was decreased significantly by treatment with mitiglinide calcium in type 2 diabetic patients,and it may play a role in the pathogenesis of type 2 diabetes mellitus.