Diagnosis, Differential ; Female ; Humans ; Infant, Newborn ; Nasopharyngeal Neoplasms ; pathology ; surgery ; Polyps ; pathology ; surgery ; Teratoma ; pathology

Objective To construct a recombinant Bacillus Calmette-Guerin ( BCG ) vaccine strain, rBCG::Rv3478-pMV261, expressing the Rv3478 protein of Mycobacterium tuberculosis and to inves-tigate its immunogenicity.Methods The gene fragments encoding Rv3478 antigen were amplified by PCR and then respectively cloned into pMV261 and pET-28a vectors to construct the recombinant expression plas-mids (Rv3478-pMV261 and Rv3478-pET-28a).The Rv3478-pMV261 plasmids were transformed into the BCG cells to construct the rBCG vaccine strains, while the Rv3478-pET-28a plasmids were transformed into Escherichia coli BL21 strains for the expression of Rv3478 protein.Polyclonal antibodies were induced in mice upon the immunization with Rv3478 protein.The rBCG vaccine strains overexpressing Rv3478 protein were screened out with Western blot assay.The C57BL/6 mice were divided into four groups including the PBS treated group, BCG treated group, rBCG::pMV261 ( R0) treated group and rBCG::Rv3478-pMV261 ( R3) treated group.All mice were sacrificed in 4 or 12 weeks after immunization.Enzyme-linked immunos-pot assay ( ELISPOT) , ELISA and flow cytometry analysis were performed to evaluate the induced humoral and cell-mediated immune responses in mice.Results The Rv3478 protein was successfully expressed and could induce polyclonal antibodies in mice.High levels of IFN-γand TNF-αwere detected in mice treated with R3, indicating that the immunization with R3 enhanced the cellular immunity.Moreover, the ratios of CD4+to CD8+T cells and the percentages of CD44+CD62L+T cells were increased in mice upon the immuni-zation with R3.Conclusion The recombinant BCG vaccine strain overexpressing Rv3478 protein could in-duce stronger cell-mediated immune responses in mice.It might be have a great significance as a new tuber-culosis( TB) vaccine strain against TB infection in the future.

Adult ; Chondroma ; pathology ; surgery ; Esophageal Diseases ; pathology ; surgery ; Esophageal Neoplasms ; pathology ; surgery ; Hamartoma ; pathology ; surgery ; Humans ; Male

OBJECTIVETo compare the intervention effects of four traditional Chinese medicines (TCMs) with typical cold or hot property on body temperature and temperature-sensitive transient receptor potential ion channel proteins (TRPs) of rats with yeast-induced fever.

METHODThe pyrexia model was induced by injecting yeast suspension subcutaneously. Totally 108 male SD rats were randomly divided into the normal group, the model group, the Rhei Radix et Rhizoma treated group, the Coptidis Rhizoma treated group, the Euodiae Fructus treated group, and the Alpiniae Officinarum Rhizoma treated group, with 18 rats in each group. At the 4 h, 8 h and 12 h after injection of yeast, the rats were sacrificed to collect their hypothalamus and dorsal root ganglion. The expressions of TRPV1 and TRPM8 were detected by immunohistochemistry and Western blot method.

RESULTCompared with the normal group, after injection of yeast, the temperature of rats in the model group notably increased, and reached the peak at 8 h (P < 0.01). The TRPV1 level in hypothalamus and dorsal root ganglia (DRG) of the model group significantly increased, whereas the TRPM8 level significantly reduced. Compared with the model group, the Rhei Radix et Rhizoma group and the Coptidis Rhizoma group showed significant decrease in the high body temperature of rats caused by yeast, down-regulation in the expression of TRPV1, and up-regulation in the expression of TRPM8 (P < 0.05 or P < 0.01). Euodiae Fructus and Alpiniae Officinarum Rhizoma had no significant effect on either temperature or TRPs of fever rats.

CONCLUSIONRhei Radix et Rhizoma and Coptidis Rhizoma, both are TCMs with cold property, can reduce the temperature of fever rats induced by yeast, which may be related to their effective regulation of TRPV1 and TRPM8 in hypothalamus and DRG, while Euodiae Fructus and Alpiniae Officinarum Rhizoma had no relevant effect.

Animals ; Antipyretics ; administration & dosage ; chemistry ; Body Temperature Regulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Fever ; drug therapy ; immunology ; microbiology ; physiopathology ; Gene Expression Regulation ; drug effects ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Saccharomyces cerevisiae ; immunology ; TRPM Cation Channels ; genetics ; immunology ; TRPV Cation Channels ; genetics ; immunology

To construct knockout vectors containing ampicillin resistant gene and partial sequence of hyaluronidase gene(Hyl)so that Hyl can be knock out by transforming the plasmid into Streptococcus zoopidemics mutans.First,partial sequence of Hyl(Hyl-1)was cloned into the vector of pMD19-T by using DNA of Streptococcus zoopidemics as template,and then a knockout vector pMD19T-SA was constructed,in which Hyl-1 gene was disrupted by inserting ampicillin resistant gene(Amp)from reverse PCR.As expected,the vector was proved to be consisted of Hyl-1-Amp-Hyl-1-pMD19-T.Thereafter,DNA fragment of Hyl-1-Amp-Hyl-1 was subcloned into pBR322 vector,the resulting construct was then checked by PCR and restriction analysis for the proper configuration of the knockout vector pBR322-SA.Both of the knockout vectors were used to transform Streptococcus zoopidemics and one recombinant was obtained in result.From results of PCR and Hyl activity assay,it was indicated that in the recombinant the Hyl gene was disrupted completely.

ObjectiveTo study tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) 1525 locus polymorphism in patients with nodular thyroid disease and investigate the relation between individual gene polymorphism and thyroid diseases.Methods A total of 125 patients were diagnosed with nodular thyroid disease at the Department of Endocrinology,the First Affiliated Hospital of Inner Mongolia Medical College.Among these patients,67 cases were nodular thyroid goiter and 58 cases were nodular thyroid adenoma,54 males,71 females,and average age was 41.05 ± 14.42. Patients with nodular thyroid goiter were grouped into toxic and non-toxic and thyroid adenoma were grouped into high-functioning or non-high-functioning.A total of 100 healthy subjects.47 males,53 females,average age 42.35 ± 16.52 were as control group.According to the principle of informed consent,venous blood was collected,polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for TRAIL gene 1525 locus polymorphism determination was performed to calculate genotype of the TRAIL gene (homozygous GG,heterozygous GA,mutationhomozygous AA) and the gene frequency (G,A),and relative degree of risk(odd ratio,OR) was compared.ResultsNodular goiter group TRAIL gene 1525 locus genotype frequencies(GG:40.3％,AG:44.8％,AA:14.9％),allele frequencies(G:62.7％,A:37.3％) were compared with that of the control group(GG:17.0％,AG:65.0％,AA:18.0％; G:49.5％,A:50.5％),the differences were statistically significant(x2 =11.376,5.633,P ＜ 0.01 or ＜ 0.05 ).Adenomas group 1525 locus genotype frequencies of the TRAIL gene(GG:44.8％,AG:38.0％,AA:17.2％),allele frequencies(G:63.8％,A:36.2％) were compared with that of the control group,the differences were statistically significant(x2 =15.342,6.054,P ＜ 0.01 or ＜ 0.05).Allele frequencies of thyroid goiter group and denoma group were compared with that of the control group,OR values were 1.714 and 1.797(all P ＜ 0.05) and 95％ confidence intervals were 1.097 - 2.679 and 1.124 - 2.874.The difference of 1525 locus genotype or allele frequency distribution in toxic and non-toxic nodular group,high functioning and non-high-functioning adenomas group were not statistically significant (x2 =3.714,2.792; 1.103,2.020; all P ＞ 0.05).ConclusionTRAIL gene 1525 locus polymorphism is significantly associated with nodular thyroid disease.

Proton pump inhibitor (PPI) are one of the most commonly prescribed classes of anti acid drugs,mainly used for peptic ulcer disease,gastroesophageal reflux disease and treatment of Helicobacter pylori.Prolonged use of PPIs is associated with a variety of adverse drug reaction.Also it probably increases the risk of dementia,though its mechanism remains unclear.This article aims to explain the correlation between PPI and dementia and its possible mechanisms.

OBJECTIVETo establish a method for the prenatal diagnosis of 22q11 microdeletion syndrome.

METHODSBACs-on-Beads (BoBs) and fluorescence in situ hybridization (FISH) were performed on a fetus for whom amniotic chromosomal culturing has failed and a pair of twin fetuses suspected for 22q11 deletion syndrome.

RESULTS22q11 microdeletion was detected in all 3 fetuses by prenatal BoBs as well as FISH, with only one red signal detected at the DiGeorge/VCFS N25 site and two green signals on the 22q13.3 ARSA site.

CONCLUSIONThe combination of prenatal BoBs and FISH can provide a method for the prenatal diagnosis of 22q11 microdeletion.

Adult ; Chromosome Deletion ; Chromosomes, Human, Pair 22 ; genetics ; DiGeorge Syndrome ; diagnosis ; embryology ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo study the stereoselective glucuronidation of carvedilol (CARV) by three Chinese liver microsomes.

METHODSThe metabolites of CARV were identified by a hydrolysis reaction with beta-glucuronidase and HPLC-MS/MS. The enzyme kinetics for CARV enantiomers glucuronidation was determined by a reversed phase-high pressure liquid chromatography (RP-HPLC) assay using (S)-propafenone as internal standard after precolumn derivatization with 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosylisothiocyanate.

RESULTSTwo CARV glucuronides were found in three Chinese liver microsomes incubated with CARV. The non-linear regression analysis showed that the values of K(m) and V(max) for (S)-CARV and (R)-CARV enantiomers were (118+/-44) micromol/L, (2 500+/-833) pmol/(min.mg protein) and (24+/-7) micromol/L, (953+/-399) pmol/(min.mg protein), respectively.

CONCLUSIONThese results suggested that there was a significant (P<0.05) stereoselective glucuronidation of CARV enantiomers in three Chinese liver microsomes, which might partly explain the enantioselective pharmacokinetics of CARV.

Carbazoles ; metabolism ; China ; Glucuronic Acid ; metabolism ; Glucuronides ; metabolism ; Microsomes, Liver ; metabolism ; Propanolamines ; metabolism ; Stereoisomerism

Objective To investigate the effect of Astragalus injection combined with low molecular heparin sodium on patients with preeclampsia.Methods All total of 64 patients with preectampsia were selected from our hospital in accordance with the standards were selected and randomly divided into two groups,each group with 32 cases.The control group was treated with 25％ magnesium sulfate 25 mL + 5％glucose injection 20 mL,slow intravenous injection in 5-10 min,followed by 25％ magnesium sulfate 40 mL + 5％ glucose injection 500 mL,intravenous infusion with 1-2 g · h-1 dripping speed,the total volume of magnesium sulfate was 25 g,qd,continued treatmcnt for 1 week.The treatment group,on the basis of the control group,treated with low-molecular-weight heparin sodiun injection 4100 U,qd,subcutaneous injection;Astragalus injection 30 mL +0.9％ normal saline 250 mL,qd,intravenous drip,continued treatment for 1 week.The percentage of platelets positive for glycoprotein GP I1 / 111 α were detected by flow cytometry.The serum D-dimer values were measured by colloidal gold double antibody sandwich nethod.The serum level of estrogen (E2) was determined by enzyme-linked immunosorbent assay.Results After treatment,factors in the treatment group and the control group were as follows:the glycoprotein GP Ⅱ/Ⅲ α levels were (0.86 ± 0.39),(1.39 ± 0.41)％;serum D-dimer levels were (1.07 ±0.37),(1.49 ±0.41) mg · L-t,the serum E2 levels were(54.63 ± 8.64) and(47.82 ± 8.21) pmol · L-1,the differences between the two groups were statistically significant(all P ＜ 0.05).The pregnancy termination week in the treatment group and the control group were (32.51 ± 1.37),(31.26 ± 1.24) weeks;the neonatal body mass in the two groups were (1956.26 ± 142.36),(1824.63 ± 132.59) g.Compared with the control group,the differences were statistically significant (all P ＜ 0.05).The incidence rates of adverse drug reactions in treatment and control groups were respectively 31.25％ (10 cases/32 cases)and 40.63％ (13 cases/32 cases),there was no statistical difference between the two groups (P ＞ 0.05).Conclusion Astragalus injection combined with low molecular heparin sodium can safely and effectively improve the pregnancy outcome of preeclampsia pregnant women.