Objective: To investigate the effect of Toxoplasma gondii ME49 strain on the apoptosis of mouse placental trophoblastic cells in vitro. Methods: Mouse placental trophoblastic cells (concentration of 5×106/ml) were cultured in the different cell culture vessels. The cells were treated for 8 h with different concentrations of Toxoplasma gondii ME49 strain (the concentration of tachyzoites was 2×106/ml, 4×10 6/ml, and 8×106/ml, respectively). FCM was used to examine the apoptosis rates of the placental trophoblastic cells stained with the fluorescent dye of Annexin V-FITC/PI; fluorescence microscopy was used to observe the changes of cellular morphology, and Western blotting analysis was used to detect the protein levels of Bax and Bcl-2. Results: The trophoblastic cells infected with Toxoplasma gondii ME49 strain showed a higher apoptosis compared to the normal cells(P<0.05), and the apoptosis rates increased with the concentration of tachyzoites in the infected groups. The highest apoptosis rate was 28.37% which was found 8 h after culture with 8×106/ml tachyzoites. Fluorescence microscope observed that the apoptosis of trophoblastic cells increased with the increase of Toxoplasma gondii. Western blotting analysis showed that the relative expression levels of Bax and Bcl-2 were 1.24±0.05, 1.37±0.03, 1.78±0.04, and 1.15±0.03, 1.09±0.05, 0.97±0.01, respectively, which were significantly different from those of the control group (1.17±0.06, 1.23±0.02, P<0.05). Conclusion: Infection with Toxoplasma gondii ME49 strain can promote the apoptosis of mouse placental trophoblastic cells in vitro through up-regulating Bax expression and down-regulating Bcl-2 expression.

Objective:To study the expressions of E-cadherin(E-cad) an d CD44V6 in normal oral mucous,oral atypical hyperplasia and oral squamous cell carcinoma(OSCC).Methods:The expression of E-cad and CD44V6 in 1 7 cases of normal oral mucous,6 of oral atypical hyperplasia and 52 of OSCC was examined with immunohistochemistry. Results: E-cad expression was observed in all the cases of normal oral mucous and oral atypical hyperplasi a,and in 43/52( 82.69% ) of OSCC. CD44V6 expression was observed in all the ca ses of normal oral mucous and oral atypical hyperplasia,and in 32/52(61.54%) of OSCC.In OSCC, the expression of E-cad was decreased with the decrease of differ entiation degree(P

Objective To investigate whether there is an inhibition of the human lung cancer cell line A549 induced by the culture supernatant of Toxoplasma gondii in vitro and the mechanism of the inhibition. Methods A549 cells 5 x 10~4mL~(-1) were cultured and harvested. The cells were treated for different hours with different concentrations of Toxoplasma gondii culture supernatant (the concentrations of tachyzoites were 4×10~7mL~(-1), 8 × 10~7mL~(-1), 16 ×10~7mL~(-1) respectively). Growth inhibition rate was measured with the MTT method; Cell cycle was checked with flow cytometer. Western blot was used to detect the level of cyclinBl and cdc2 of cells. Results The culture supernatants of Toxoplasma gondii inhibited proliferation of A549 cells in a time-dose dependent manner. Cell cycle was significantly stopped at G_2/M phase by the culture supernatants with FCM technology. The culture supernatant of Toxoplasma gondii reduced the expressions of gene cyclinBl and cdc2 of A549 cells. Conclusion The culture supernatant of Toxoplasma gondii may inhibit A549 cell and arrest the cell cycle of A549 cells mainly by regulating the expression of gene cyclinBl and cdc2.

Objective To evaluate the effect of acupuncture for treating migraine acute attack to offer some evidence‐based basis for clinical application .Methods The Chinese and English literatures on the acupuncture for treating migraine acute attack were retrived from January 1989 to December 2014 ,the literatures were screened according to inclusion and exclusion criteria ,the Meta‐analysis was performed on these chose literatures .Results A total of 5 studies were included and 618 migraineurs were in‐volved ,four literatares were performed the Meta‐analysis ,and 1 literature was performed the description analysis .Meta‐analysis re‐sults showed that there was statistically significant differences between the acupuncture group and the sham acupuncture group in the VAS score reduction value at 2 h[MD=0 .36 ,95% CI:0 .08 ,0 .65 ,P=0 .01] ,4 h[MD=0 .49 ,95% CI:0 .14 ,0 .84 ,P=0 .007] after acupuncture;while when the VAS score was used as the evaluation indicator ,there was no statistically significant differences were found at 2 h[MD= -0 .38 ,95% CI:-0 .83 ,0 .07 ,P=0 .10] ,4 h[MD= -0 .42 ,95% CI:-0 .96 ,0 .12 ,P=0 .12] after acu‐puncture in the VAS score between the acupuncture group and the sham acupuncture group .Conclusion Acupuncture could effec‐tively relieve the intensity of headache in migraine ,the analgesic effect of acupuncture for treating migraine attacks is significantly superior to the sham acupuncture group ,while with the VAS score as the evaluation indicator ,the difference between the acupunc‐ture group and the sham acupuncture group has no statistical significance .

The partial segment of Marek′s disease virus (MDV) glycoprotein B (gB) gene was amplified by PCR. The segment was cloned into pET-28a vector to obtain the recombinant pET-gB plasmid. The recombinant plasmid was transformed into E.coli BL21,and expressed in very high level as inclusion body after induced with 1.0mmol/L IPTG. The inclusion body was solubilized in urea (8mol/L) . The purified protein was obtained by use of His?Bind affinity chromatography. Mice were immunized i.p. by the purified protein to make the polyclonal antibody. The titer of the antibody by indirect ELISA was 1?10~ -5 . Moreover, the analysis by western blot proved that antibody was specific to the recombinant protein. These works lay a favorable foundation for the study of the immune response by MDV gB.

Objective Toevaluatea modified Ziehl-Neelsen(Z-N) stain in the diagnosis of tuberculous meningitis. Methods Cerebrospinal fluid specimens from 35 patients were stained by using the modified Ziehl-Neelsen staining. Re-sults The positive rate was 94.29% in 35 patients with tuberculous meningitisand the intracellular acid-fast bacilli was detected in 53.40%of all specimens. One case was stained positive in 15 patients with non-tuberculous meningitis. Con-clusion The modified Ziehl-Neelsen stain not only significantly improves the detection rates of tuberculous meningitisbut alsois able to identify intracellular M.tuberculosisin cerebrospinal fluidspecimen.Thus, the modified Z-N stain can be a convenient tool for diagnosing tuberculous meningitis.

The expressing cassette, LoxP-CMV-gpt-IRES-LoxP( about 2.9kb), was amplified by PCR from a plasmid, pIRES-gpt, by use of the primers , which contained the loxP sites in 5' terminals, respectively. The loxP sites were designed into primers by the software of Primer primer 5.0. Then the cassette was cloned into the site of BalI in pBUS10 to obtain pUS-gptIRES(L). The sequencing analysis for pUS-gptIRES(L) indicated that two loxP sites with the same direction were correctly inserted into pUS-gptIRES(L).The gpt gene in pUS-gptIRES(L) was replaced by a fragment including the full length GFP gene as well as SV40 poly A sequence to get pUS-GFPIRES(L). pUS-GFPIRES(L) was transiently transfected into CHO cell lines, and then the green fluorescence could be seen, the results showed that GFP gene could be expressed correctly. Moreover, pUS-GFPIRES(L) was transfected into the CEF infected MDV CVI988 strain and recombinant virus was selected by the green fluorescence. The growth curve of virus showed the characteristic of recombinant virus was the same as that of CVI988 in vitro. These results give the basis for further studying the characteristic of MDV in vivo and the application of the Cre/LoxP system to MDV genome.

OBJECTIVETo explore the relationship between quinone oxidoreductase1 (NQO1) gene nonsynonymous cSNP and the genetic susceptibility of esophageal cancer.

METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and Allele-Specific PCR (AS-PCR) were employed to assess the polymorphism of NQO1 genes both in 106 patients with esophageal cancer and control subjects matched by age, gender and origin.

RESULTSIt was shown that no C/C genotype was found at 406 of NQO1. The allelic frequency of NQO1 609T was significantly higher in patients with esophageal cancer than in the control subjects (P < 0.005) and the individuals with 609T allelic genotype of NQO1 gene were at greater risk to develop esophageal cancer (OR = 4.76, 95% CI = 1.064 - 3.397). But Individuals with mutant allele of NQO1 465 genotype did not show the rising risk of esophageal cancer.

CONCLUSIONSThe NQO1 C609T polymorphisms should likely be associated with the genetic susceptibility of esophageal cancer.

Alleles ; China ; Esophageal Neoplasms ; ethnology ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; NAD(P)H Dehydrogenase (Quinone) ; genetics ; Polymorphism, Single Nucleotide

Objective To investigate expressions of phosphorylated c-Jun N-terminal kinase (p-JNK)and P38 mitogen-activated protein kinase(p-P38MAPK)in psoriasis vulgaris lesions. Methods Tissue specimens were obtained from lesions of 30 patients with psoriasis vulgaris and normal skin of 30 healthy human controls. An immunohistochemical study and Western-blot analysis were performed to measure protein expressions of p-JNK and p-P38MAPK in these skin specimens. Results As the immunohistochemical study showed, the expressions of p-JNK and p-P38MAPK(expressed as the average optical density [AOD]value for targeted proteins)were significantly higher in psoriasis vulgaris lesions than in normal skin tissues (p-JNK: 0.663 ± 0.016 vs. 0.333 ± 0.009, t = 44.869, P < 0.001; p-P38MAPK: 0.436 ± 0.011 vs. 0.306 ± 0.010, t = 21.913, P < 0.001). Western-blot analysis also showed increased protein expressions of p-JNK and p-P38MAPK in psoriasis vulgaris lesions compared with normal skin tissues (t = 20.477, 165.084, respectively, both P <0.05). Conclusion The activation of JNK and P38MAPK may be involved in the overproliferation of epidermal cells in psoriasis vulgaris lesions.

Objective To investigate the effect of combined use of insulin and acarbose on glucose excursion in type 1 diabetic patients.Methods 120 cases were randomly divided into control group and observation group.The control group received preprandial ultra-short effect insulin and long-acting insulin before bedtime while the observation group received acarbose 50 mg added to the medicine taken by the control group.Continuous Glucose Monitoring System (CGMS) was used to watch the blood glucose fluctuations.Data related to blood glucose level,glucose excursions after meals and hypoglycemia at night were compared between patients in the two groups.Results The average blood glucose (9.37 ± 1.70) mmol/L,the largest amplitude of glycemic excursions (LAGE) ( 11.42 ± 2.73 ) mmol/L,hyperglycemia-area under curve 0.89 ± 0.54,mean amplitude of glycemic excursions (MAGE) (5.13 ± 2.23) mmol/L,M-value (18.93 ± 11.43) mmol/L and insulin dosage (42.11 ± 14.42)U/day of observation group were significantly lower than in the control group (P＜0.05 ).Glucose excursions after meals and the times( 0.33 ± 0.50 )/day,the maintenance time (43.75 ± 43.50)/min and low glycemic index ( LBGI ) (0.005 ± 0.002 ) mmol/L of hypoglycemia at night were also significantly lower than in the control group,with statistically significant (P＜0.05) differences.Conclusion The blood glucose fluctuation was significantly improved,with the decrease of insulin dosage while both glucose excursions and hypoglycemia at night reduced in patients with typel diabetes mellitus after the acarbose treatment.We suggested that this program deserve further observation.