Objective To investigate the effect of exogenous hydrogen sulfide (H 2 S) on H2 S concentration and cystathionine β-synthase (CBS) expression in hippocampus in a rat model of vascular dementia (VaD). Methods A rat model of VaD was induced by using the modified four -vessel occlusion. The rats were divided into sham operation, model, low -dose and high-dose NaHS groups using the random number table method. They were further redivided into one day, seven -day, and 30-day subgroups according to the time after modeling. After modeling respectively, NaHS 30 μmol/kg and 100 μmol/kg were injected intraperitoneally every day in the low -dose and high-dose NaHS groups. The normal saline was injected intraperitoneally every day in the sham operation group and the VaD model group. Morris water maze test was used to evaluate the learning and memory ability of the rats. The expression of CBS in hippocampus was detected by real-time fluorescent polymerase chain reaction. Western Blotting was used to detect expression of CBS protein in hippocampus. Results Morris water maze test showed that the escape latencies of the model group, low -dose and high-dose NaHS groups were prolonged significantly compared with the sham operation group (P <0.05); the times of crossing the platform were decreased significantly compared with the model group (P <0.05); and the escape latencies were shortened significantly in the low -dose and high-dose NaHS groups compared with the model group ( P <0.05). The H2 S content in hippocampus was decreased significantly in the model group, low -dose and high-dose NaHS groups compared with the sham operation group, but the low -dose and high-dose NaHS group was significantly higher than that in the model group (all P <0.05). The expression of CBS mRNA and protein in the model, low -dose and high-dose NaHS groups was significantly lower than that of the sham operation group (all P < 0.05), and there was no significant difference between the low -dose and high-dose NaHS groups and the model group. Conclusions Exogenous H2 S may improve the learning and memory ability of the VaD rats. It may be associated with the increased H2 S content in hippocampus. However, it has no effect on CBS expression.

OBJECTIVE: To investigate the genetic polymorphisms of 19 STR Loci in Shandong Han population in order to provide the genetic data for paternity testing. METHODS: The genotypes of 205 unrelated individuals in Shandong Han population were typed by Goldeneye 20A kit to get the allele frequencies and population genetic parameters of 19 STR loci. Four kits, Identifiler kit, SinoFiler kit, PowerPlex 16 kit, and Goldeneye 20A kit, were compared with each other and used in the analysis of a special paternity test case. RESULTS: The population genetic parameters of 19 STR loci in Shandong Han Population were obtained. The cumulative discrimination power (CDP) and cumulative probability of exclusion (CPE) ranked from high to low were Goldeneye 20A kit, SinoFiler kit, PowerPlex 16 kit and Identifiler kit, respectively. As duo case, the result of the real case showed that Identifiler kit had no excluding loci, and none of the SinoFiler kit, PowerPlex 16 kit or Goldeneye 20A kit could exclude fatherhood. CONCLUSION Compared with Identifiler kit, SinoFiler kit, and PowerPlex 16 kit, Goldeneye 20A kit shows the higher efficiency than the others, but is not completely satisfied for duo cases.
Asian People/genetics* ; China ; Forensic Genetics/methods* ; Gene Frequency ; Genetic Loci/genetics* ; Genetics, Population ; Genotype ; Humans ; Male ; Microsatellite Repeats ; Paternity ; Polymorphism, Genetic/genetics*

The beta-glucosidase encoding gene bglA was cloned from Bacillus polymyxa 1.794. The bglA gene was inserted in expression vector pET28a(+) and transformed into Escherichia coli BL21 (DE3), finally the recombinant strain BL1979 was obtained. Induced by IPTG, the expression P-glucosidase activity reached to 24.7 IU/mL. The optimum temperature and optimum pH of the recombinant expression P-glucosidase in BL1979 were 37 degrees C and 7.0 respectively,the purity can reach to 92.7%. Analysis of the fusion protein by nondenaturing gradient gel electrophoresis, we found the fusion protein exists in dimmer, tetramer,hexamer and octamer, they all have hydrolase activity.
Bacillus ; enzymology ; Escherichia coli ; genetics ; Plasmids ; Recombinant Proteins ; biosynthesis ; isolation & purification ; beta-Glucosidase ; genetics ; isolation & purification ; metabolism

OBJECTIVETo study the cell biocompatibility of porous biphasic calcium phosphate nanocomposite in vitro.

METHODSBone marrow mesenchymal cell (BMSCs) obtained from SD rat bone marrow were in vitro induced and proliferated. Afler their osteoblast phenotypes were verified, BMSCs were seeded onto prepared porous biphasic calcium phosphate nanocomposite (Experiment group) and common porous hydroxyapatite (Control group). The cell adhesion was evaluated by scanning electron microscope. Synthesis of alkaline phosphatase enzyme (ALP) and osteocalcin were detected and cell cycle was detected by flow cytometry.

RESULTSBMSCs could fully attach to and extend on the material in experiment and control group, Moreover, experiment group were superior to control group in adhesion, proliferative abilities and osteogenic activity.

CONCLUSIONBMSCs can differentiate to osteoblast phenotype; the porous biphasic calcium phosphate nanocomposite as bone tissue engineering scaffold has good cell biocompatibility.

Alkaline Phosphatase ; Animals ; Bone Marrow Cells ; Bone and Bones ; Cell Adhesion ; Durapatite ; Hydroxyapatites ; Materials Testing ; Nanocomposites ; Osteoblasts ; Osteocalcin ; Rats ; Tissue Engineering ; Tissue Scaffolds

Haploinsuffieiency of the runt-related transcription factor 2 (Runx2) gene is widely known to be responsible for cleidocranial dysplasia (CCD).To date,more than 190 mutations in Runx2 gene have been reported to be related to CCD.In this study,a novel mutation of Runx2 gene was observed in a female with CCD.Genomic DNA was extracted from peripheral venous blood of the proband and eleven members of her family.Genetic testing on these twelve people identified a novel missense mutation (c.895T＞C,Y299H) in exon 5 of the RUNX2 gene in the proband.This mutation results in an amino acid change at codon 895 (P.Tyr 299 His.) from a tryptophan codon (TAT) to a histidine codon (CAT).Our finding may further extend the known mutation spectrum of the RUNX2 gene,and facilitate prenatal genetic diagnosis of CCD in the future.

Objective:To explore the expression pattern of pyroptosis-related genes(PRGs)in hepatocellular carcinoma(HCC),and analyze the relationship between its expression and tumor prog-nosis and immune microenvironment.Methods:TCGA database was used to analyze the genetic changes and expression patterns of PRGs in primary HCC cells,and cluster analysis was used to i-dentify the pyrogenic subtypes of HCC.To compare the difference of prognosis and immune mi-croenvironment among HCC pyrodeath subtypes.Scorch death score quantified the comprehensive expression of PRGs in each sample,and analyzed the correlation between scorch death score and each immune score.Results:Two pyroptosis-associated subtypes of primary HCC were identi-fied,and the expression pattern of PRGs is closely related to the prognosis of cancer patients and the tumor microenvironment.The subtype with high expression of PRGs had a poor prognosis,and functional enrichment analysis found that some tumor-promoting pathways and PD-1 checkpoint pathways were significantly enriched in this subtype.And various cells and immune checkpoints re-lated to immunosuppression were also enriched in this subtype.By constructing PYROPTO-SIS_score to quantify the comprehensive expression of pyroptosis-related genes in each sample,it was found that PYROPTOSIS_score was significantly positively correlated with tumor-infiltrating macrophages,myeloid-derived suppressor cells,and Treg cells.Conclusion:These results sug-gest that pyroptosis may play a tumor-promoting as well as immunosuppressive role in HCC,pro-viding new insights into the assessment of tumor patient prognosis and the immune microenviron-ment.

AIMTo evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI).

METHODSFifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration.

RESULTSA total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term.

CONCLUSIONIt appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.

Adult ; Cell Culture Techniques ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Female ; therapy ; Infertility, Male ; therapy ; Male ; Oocytes ; growth & development ; Pregnancy ; Pregnancy Rate ; Semen ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; Testis ; cytology

OBJECTIVETo investigate the clinical value of artificial insemination by donor (AID).

METHODSWe retrospectively analyzed 480 cycles of AID among 258 infertile couples, who were divided according to the women's age into a < or = 30 yr group and a > or = 31 yr group.

RESULTSA total of 120 pregnancies were achieved in 480 AID cycles, with a cycle pregnancy rate of 25.00% and a cumulative pregnancy rate of 46.51%. In the natural cycles, the cycle pregnancy rate was 29.65% and the cumulative pregnancy rate was 51.00% in the < or = 30 yr group, significantly higher than 13.33% and 25.00% in the > or = 31 yr group (P < 0.05). In the ovulation induction cycles, no significant differences were found in the cycle and cumulative pregnancy rates between the two groups (24.02 and 48.86% versus 23.81 and 43.48% , P > 0.05). The cycle and cumulative pregnancy rates decreased with the increase of infertility duration and the women's age, but had no significant differences. In the first four cycles, the cycle pregnancy rates were 24.03, 24.94, 24.69 and 25.00% (P > 0.05), and the cumulative pregnancy rates were 24.03, 39.53, 45.74 and 46.51%, with significant differences between the first cycle and the other three (P < 0.01).

CONCLUSIONOvulation induction is superior to natural cycle in AID for older women. IVF/ICSI can be resorted to only after AID has failed three or four times.

Adult ; Azoospermia ; therapy ; Female ; Humans ; Insemination, Artificial, Heterologous ; Male ; Ovulation Induction ; Pregnancy ; Pregnancy Rate ; Retrospective Studies

OBJECTIVETo investigate the roles of the residential environment and eating habits in the pathogenesis of bronchial asthma in school children.

METHODSOne hundred and twenty-nine children between 6-12 years who were diagnosed with asthma were enrolled. Two hundred and fifty-eight healthy age- and gender-matched children were used as the control group. A questionaire which included 23 factors related to respiratory tract anaphylactic diseases such as residential environment and eating habits were completed by the children's parents.

RESULTSLogistic regression analysis showed that 6 variances out of 16 agents of the residential environment, the experience of raising pets, the type of floor, the type of pillow, the type of quilts, the heating equipments and the house area, were entered into the regression equation; none of the 7 variances of eating inhabits was entered into it.

CONCLUSIONSThe residential environment plays an impotent role in the pathogenesis of bronchial asthma in children. The incidence of bronchial asthma in children can be reduced by the improvement of the residential environment.

Asthma ; etiology ; Case-Control Studies ; Child ; Female ; Humans ; Logistic Models ; Male ; Risk Factors

OBJECTIVETo detect the alterations of mitochondrial 12S rRNA in patients with gastric cancer, and further evaluate their effects on development of gastric carcinomas.

METHODSMitochondrial 12S rRNA of 22 samples of gastric cancer tissues and 22 corresponding normal gastric mucosa taken from the distal portion of surgical specimens were PCR amplified, followed by direct DNA sequencing. Laser capture microdissection technique (LCM) was used to isolate cancerous cells and dysplastic cells from patients with specific mutations. Denaturing high-performance liquid chromatography (DHPLC) plus allele-specific PCR (AS-PCR), nest-PCR and polyacrylamide gel electrophoresis (PAGE) were applied to further evaluate this mutant property and quantitative difference of mutant type between cancerous and dysplastic cells. Finally, RNAdraw bio-soft was used to analyze the RNA secondary structure of mutant type 12S rRNA.

RESULTSCompared with mitomap database, some variations were firstly found, among which np652 G insertion and np716 T-G transversion were only found in cancers. There existed statistically significant difference in variant frequency of 12S rRNA between intestinal type and diffuse type of gastric carcinoma, 5/17 (29.4%) and 12/17 (70.6%) respectively (P < 0.05). DHPLC analysis showed that 12S rRNA np652 G insertion and np716 T-G transversion were heteroplasmic mutation. Variant frequency of 12S rRNA in cancer was higher than that in dysplasia (P < 0.01). 12S rRNA 652G insertion had more adverse effect on secondary structure stability of 12S rRNA than T-G transversion did.

CONCLUSIONHighly variant frequency of mitochondrial 12S rRNA may be associated with intestinal type of gastric cancer. Most parts of variations exist in both cancer and normal tissues and may not be characteristic of tumor specificity. However np652 G insertion and np716 T-G transversion may possess some molecular significance on gastric cancerogenesis. During the process of progression from normality through dysplasia to cancer, 12S rRNA tended to transit from homoplasmy (wild type) and heteroplasmy to homoplasmy (mutant type, np717 T-G).

Base Sequence ; Chromatography, High Pressure Liquid ; methods ; Humans ; Molecular Sequence Data ; Point Mutation ; RNA ; genetics ; RNA, Ribosomal ; genetics ; Stomach Neoplasms ; genetics ; Tumor Cells, Cultured