Asthma ; genetics ; Child ; Child, Preschool ; Gene Frequency ; Genotype ; Humans ; Infant ; Male ; Peptidyl-Dipeptidase A ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic

Objective To evaluate the dynamic contrast-enhanced MRI (DCE-MRI)in detecting experimentally induced testicular ischemia. Methods Thirty healthy male New Zealand rabbits were randomly assigned into 6 groups. There were 5 rabbits in each of the following experimental groups: ( 1 ) Normal control, (2) Sham-operated, (3) ischemia of 3 h group, (4) ischemia of 6 h group, (5) ischemia of 12 h group, (6) ischemia of 24 h group. In all experiment groups, the right testis served as the internal control while the left testis served as the experimental side. DCE-MRI for each animal lasts about 10 minutes. Signal enhanced ratios (SERs) of ROI for both sides of each group were calculated by a computer, and parameters of SERs of 30 s, 75 s, 120 s and maximal SER were used for statistical analysis.Time intensity curves (TICs) were made for two sides of each group via Excel 2003 software and classified into 4 types. Statistical analysis was performed to compare the differences of SERs between left and right testis by two independent Kolmogorov-Smirnov test. Results In group I and 2, significant enhancement was observed on both testes of 10 rabbits. The enhancement decreased gradually with the elongation of ischemia in torsion groups. Three cases of type Ⅰ and 2 cases of type Ⅱ were observed in group 1,5 cases of type Ⅰ in group two, 2 cases of type Ⅰ and 3 cases of type Ⅱ b in group three, 2 cases of type Ⅰ and 2 cases of type Ⅱ b in group four, 5 cases of type Ⅱ b in group five and 5 cases of type Ⅲ in group six were noticed in the left testes. And in TICs of right testes, all cases showed TICs of type Ⅰ except 2 cases of type Ⅱ a in group six. In four torsion groups, the values for SER75 of the left side were 0. 084%, 0. 076%, 0.164% and 0.065%, while the right side were 0.255%, 0.410%, 0.586% and 0.302% (P ＜0.05). The values for SER120 in group three, five and six were 0.221% , 0.158% and 0.059% for the left side, and 0.405%,0.522% and 0.207% for the right side(P ＜0.05). The values for MSER in group three, five and six were 0.217% ,0.164% and 0.072% for the left side, and 0.405%, 0.586% and 0.302% for the right side(P ＜0.05). Conclusion DCE-MRI technique may be useful in the diagnosis of testicular torsion, which shows potential in the clinical application.

OBJECTIVETo investigate the expression of a novel metastasis-inducing protein human anterior gradient-2 (AGR2) in breast cancer and its clinical and prognostic significance.

METHODSAGR2 expression was assessed in 160 cases of breast cancer and 20 cases of benign breast diseases by immunohistochemistry using tissue chip technology. In addition the expression of ERa, PR and c-erbB-2 in breast cancer was also evaluated. Follow-up information of 5-year duration was available in 127 patients with breast cancer. Kaplan-Meier analysis and COX regression model were used to analyze the correlation between AGR2 expression and the follow-up clinical data.

RESULTSThe expression of AGR2 was significantly higher in breast cancers than that in benign diseases (68.3% vs. 25.0% , P < 0.01). There was a negative correlation between AGR2 expression and the histological grade of breast cancer (P <0.05) , whereas positive correlations was found between the expression of AGR2 and ERalpha (P <0.05), and between the expression of AGR2 and PR (P <0.01). In the subgroup of ERalpha-positive breast cancer, Logistic regression model demonstrated AGR2 and TNM stage were important factors affecting lymph node metastasis (both P < 0.01). Kaplan-Meier analysis demonstrated that a positive expression of AGR2 was associated with poor overall survival and relapse-free survival (both P <0.01). Moreover, COX regression model confirmed the expression of AGR2 as an independent prognostic factor among patients with ERa-positive breast cancer (P <0.01).

CONCLUSIONSThe abnormal expression of AGR2 may play a role in the pathogenesis and progression of breast cancer. The metastasis-inducing capability of AGR2 may be partly regulated through the ER pathway. Therefore, AGR2 may be a useful molecular marker for prognostication for patient with hormone-responsive breast cancer.

Antineoplastic Agents, Hormonal ; analysis ; BRCA2 Protein ; genetics ; metabolism ; Biomarkers, Tumor ; analysis ; Breast Neoplasms ; diagnosis ; genetics ; metabolism ; Estrogen Receptor alpha ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; Humans ; Immunohistochemistry ; Neoplasm Metastasis ; diagnosis ; Neoplasm Staging ; Prognosis ; Proteins ; genetics ; metabolism ; Receptor, ErbB-2 ; analysis ; metabolism

OBJECTIVETo culture and amplify the young rabbit's bone marrow stromal cells (BMSCs) in vitro, and to observe the effect of hypothermia on the cells' growing behavior and biological function.

METHODSBMSCs were acquired from the rabbit' tibia bone marrow and induced to mature osteoblasts in vitro. The cultured cells growing well in vitro were preserved in liquid nitrogen. The anabiotic cells having cryopreserved for 1 week were chosen as the experimental group, and the routine 7th generation as the control group. Their biological function in comparion by the examination of morphological changes, cells' proliferation ability, colone forming ratio, synthesis ability of ALP and protein, mineralized nodes forming ability were observed.

RESULTSAs contrast to the control groups, the anabiotic cells also grew and proliferated well in vitro except a little more slowly than before. They had the similar general shape in all the time segments, but a little differences in cells' ultrastructure. The experimental groups also had the typical characters of mature osteoblasts, and high abilities of the synthesis of ALP and proteins. The statistic data showed that these two groups had no significant difference (P > 0.05).

CONCLUSIONThe cryopreserved osteoblasts had the same biological functions and the similar growing behaviors as before. These results suggest that it is practical to use the cryopreserved osteoblasts for further study on bone tissue engineering.

Animals ; Bone Marrow Cells ; Bone and Bones ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; In Vitro Techniques ; Osteoblasts ; Rabbits ; Tissue Engineering

To establish a method for detecting microdialysis recovery of tetramethylpyrazine (TMP) and ferulic acid (FA) and investigating the influencing factors, providing the basis for further in vivo microdialysis experiments. The concentration of FA and TMP in dialysates were determined by high pressure liquid chromatography ( HPLC) and probe recovery were calculated respectively. The influence of the flow rates, medium concentration, temperature and in vivo probe stability on the recovery of FA and TMP were investigated by using concentration difference method (incremental method and decrement method). The recovery obtained by incremental method were similar to by decrement method. The in vitro recovery rate of FA and TMP decreased with the increase of 1-2.5 μL min(-1), and increased obviously with the temperature of 25-42 degrees C under the same conditions. The concentration of FA and TMP had no obvious effect on the probe recovery under the same flow rate. In addition, the recovery of TMP and FA remained stable and showed similar trends under the condition of four concentration cycles, indicating that the intra day reproducibility of the concentration difference method was good. The recovery of brain microdialysis probes in vivo 8 h maintained a relatively stable, but certain differences existed between different brain microdialysis probes, demonstrating that each probe was required for recovery correction in vivo experiment. Microdialysis sampling can be used for the local brain pharmacokinetic study of FA and TMP, and retrodialysis method can be used in probe recovery of FA and TMP in vivo.
Animals ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; Coumaric Acids ; analysis ; isolation & purification ; pharmacokinetics ; Drugs, Chinese Herbal ; Humans ; Microdialysis ; methods ; Pyrazines ; analysis ; isolation & purification ; pharmacokinetics ; Rats

OBJECTIVETo investigate the clinical phenotype and genetic characteristics of an azoospermia patient with ring 22 chromosome syndrome.

METHODSWe analyzed the clinical data of an azoospermia patient with ring 22 chromosome syndrome and reviewed relevant literature.

RESULTSThe patient was a short 29-year-old male, with bilateral testes small in size and soft in texture. Seminal examination indicated azoospermia. Chromosome analysis showed the karyotype of the patient to be 46, XY, r (22) (p11, q25). The level of testosterone was low, and the testicular tissue was brittle and easy to break. Pathological microscopy revealed reduced number of Sertoli cells and germ cells in the seminiferous tubules and thinner layers of cells. All the germ cells were spermatogonia. Neither spermatocytes nor sperm cells were found, which suggested complete spermatogenic failure. Mild interstitial fibrosis was visible in part of the seminiferous tubule walls.

CONCLUSIONPatients with ring 22 chromosome syndrome usually represent normal clinical phenotypes. However, this kind of genetic abnormality often induces severe testicular damage and spermatogenic arrest, which may result in azoospermia.

Adult ; Azoospermia ; etiology ; genetics ; Chromosomes, Human, Pair 22 ; Humans ; Male ; Oligospermia ; Ring Chromosomes ; Spermatogenesis ; Spermatogonia ; Syndrome

AIMThe process of vascular calcification involves various genetic alterations which may play a very important role in the vascular calcification. Vascular smooth muscle cells undoubtedly composed the main part of vascular cells, and are involved in vascular calcification. So bovine artery smooth muscle cell (BASMC) was used to investigate the gene changes during BASMC's calcification.

METHODSBovine artery smooth muscle cells cultured in vitro was induced calcified by beta-Glycerophosphate (beta-GP). Using DD-PCR technique to screening differentially expressed genes and those differentially expressed bands were reexamined by reverse Northern blot. All the ESTs were sequenced and BLAST with GenBank.

RESULTSTotal 65 cDNAs were isolated as differentially expressed genes and 40 of them were successfully reamplified. Using reverse-Northern blot, seven of these 40 cDNAs were reproducibly expressed differentially between the two cells. Three of them are new bands and have not been reported before.

CONCLUSIONThis is the first time using DD-PCR to screen differentially expressed genes of BASMC calcification. Seven related ESTs were identified relating to BASMC calcification.

Animals ; Arteriosclerosis ; genetics ; metabolism ; pathology ; Cattle ; Cells, Cultured ; Expressed Sequence Tags ; Genetic Variation ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; pathology ; Vascular Calcification ; genetics ; metabolism ; pathology

Objective To analyze the property of the new antibacterial agent (N, N-di-n-decyl-N, N-dimethyl-ammonium 5-oxopyrrolidine-2-carboxylate, DAPC) to prevent Streptococcus mutans' adhesion on tooth surface.Method determine the minimal inhibitory concentration and minimal bactericidal concentration of DAPC and Chlorhexidine gluconate by liquid dilution method.Set the Chlorhexidine gluconate as the positive control, while PBS as the negative control.Use the crystal violet staining to measure the quantity of biofilm.Re s ult The MIC of DAPC on Streptococcus mutans was 0.0031250％ and MBC was 0.0062500％. The MIC of Chlorhexidine gluconate was 0.0015625％, while the MBC was 0.0031250％.When concentration of the two antibacterial agents was MIC, the quantity of biofilm have no significance among three groups.However, when concentration of Chlorhexidine gluconate and DAPC were 0.12％ and 0.24％ respectively, biofilm of experimental group was lower than PBS, and there is no significance between Chlorhexidine gluconate group and DAPC group after 24 hours incubation.Conclus ion New antibacterial agent DAPC have significant effect in inhibiting of Streptococcus mutans.And the residual of DAPC on teeth can maintain a long time of antibacterial effect to inhibit Streptococcus mutans' adhesion.

Liver organoids are three-dimensional cellular tissue models in which cells interact to form unique structures in culture. During the past 10 years, liver organoids with various cellular compositions, structural features, and functional properties have been described. Methods to create these advanced human cell models range from simple tissue culture techniques to complex bioengineering approaches. Liver organoid culture platforms have been used in various research fields, from modeling liver diseases to regenerative therapy. This review discusses how liver organoids are used to model disease, including hereditary liver diseases, primary liver cancer, viral hepatitis, and nonalcoholic fatty liver disease. Specifically, we focus on studies that used either of two widely adopted approaches: differentiation from pluripotent stem cells or epithelial organoids cultured from patient tissues. These approaches have enabled the generation of advanced human liver models and, more importantly, the establishment of patient-tailored models for evaluating disease phenotypes and therapeutic responses at the individual level.

OBJECTIVETo explore F (13) A gene mutation in a pedigree with hereditary coagulation factor XIII (FXIII) deficiency.

METHODSThe FXIII deficiency was diagnosed by clot solubility test and other standard laboratory clotting tests. All exons, exon-intron boundary sequences of F(13) A gene were amplified by PCR and the products were sequenced directly. Any mutation identified by direct sequencing was confirmed by reverse sequencing. The mutation identified in the proband was screened in the family members.

RESULTSThe assays of PT, Qiulan, fibrinogen leveling, platelet counts, bleeding time were normal and the clot solubility test was positive in the proband. The homozygous deletion of 33 nucleotides (127067de133) in exon 10 of F(13) A gene which resulted in deletion of 11 amino acids in FXIIII A protein with 720aa residues was identified in the proband. Family studies showed that the mutation was inherited from the parents both of whom carried the heterozygous deletion mutation.

CONCLUSIONThe homozygous 127067de133 mutation of F(13) A gene is responsible for the disorder of the pedigree.

Adolescent ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Heterozygote ; Homozygote ; Humans ; Male ; Pedigree ; Sequence Deletion