Association of the interleukin-10 gene -1082A/G genetic polymorphism with risk of ischemic stroke in a Chinese population.

We investigated the possible association between two single nucleotide polymorphisms of IL10 (-1082A/G and -592C/A) and susceptibility to ischemic stroke. In total, 335 patients with proven ischemic stroke and 335 control subjects were recruited from Xinxiang Central Hospital between March 2013 and May 2015. The IL10 -1082A/G and -529C/A polymorphisms were investigated by polymerase chain reaction-restriction fragment length polymorphism. When compared with the control subjects, patients with ischemic stroke were more likely to be male, have a habit of tobacco smoking, have higher BMI, have hypertension or diabetes mellitus, and have higher levels of TC, LDL-C, HDL-C, and TG. The multivariate logistic regression analyses revealed that the AA genotype of IL10 -1082A/G was significantly associated with development of ischemic stroke in a Chinese population compared with the GG genotype (OR = 1.93, 95%CI = 1.15-3.25). In the dominant model, the association between GA+AA genotype of IL10 -1082A/G and risk of ischemic stroke was also significant compared with the GG genotype, and the adjusted OR (95%CI) for the GA+AA genotype was 1.41 (1.02-1.94). Thus, our study suggests that IL10 gene polymorphisms contribute to the development of ischemic stroke.

High MMP-1, MMP-2, and MMP-9 protein levels in osteoarthritis.

Our study examined the relationship between the expression of matrix metalloproteinases (MMP)-1, MMP-2, and MMP-9 proteins and the pathogenesis of osteoarthritis (OA). We employed rigorous inclusion and exclusion criteria in computer-based bibliographic databases to extract published studies relevant to this investigation. The STATA 12.0 software was used for the statistical analyses. A total of 1408 studies were initially searched, and 10 studies with 458 OA patients and 295 healthy controls were included in this meta-analysis. The meta-analysis results suggested that the protein levels of MMP-1, MMP-2, and MMP-9 were higher in patients with OA than those in the control group. A subgroup analysis according to ethnicity showed that the protein levels of MMP-1 and MMP-2 were higher in Asian patients with OA than in controls. Caucasians showed no statistically significant differences in protein expression of MMP-1 and MMP-2 between the OA patient group and the control group. Interestingly, the protein levels of MMP-9 in patients with OA were higher than those in the control group in both Asians and Caucasians. A sample-source analysis suggested that the serum levels of MMP-2 and MMP-9 proteins were higher in patients with OA than in controls, while MMP-1 and MMP-9 protein expressions were higher in the synovial joint fluid of patients with OA than in controls. In conclusion, our meta-analysis results suggested that the increased expression of MMP-1, MMP-2, and MMP-9 proteins might be associated with the pathogenesis of OA.

Association of the VRK2 gene rs3732136 polymorphism with schizophrenia in a Northwest Chinese Han population.

Previous studies have found that the vaccinia related kinase 2 gene (VRK2) polymorphism was associated with schizophrenia (SCZ) in the worldwide population. This association was further supported by VRK2 mRNA expression patterns and brain structure variations. Here, we analyzed four single nucleotide polymorphisms (SNPs) of the VRK2 gene in a total population of 893 samples, consisting of 360 patients with SCZ and 533 healthy controls of Han Chinese descent using the SNPscan method. Single SNP, haplotype, and gender-specific association analyses were performed. We found that rs3732136 was significantly associated with SCZ (P = 0.042; odds ratio = 1.25; 95% confidence interval = 1.01-1.55). Further genotype and haplotype association analyses suggested a similar pattern. Our data provide preliminary evidence that the VRK2 gene might play a major role in the development of SCZ in the Northwest Chinese Han population.

Codon optimization enhances the expression of porcine ß-defensin-2 in Escherichia coli.

Porcine ß-defensin-2 (pBD2) is a cationic antimicrobial peptide that has therapeutic potential. The amount of pBD2 in nature is limited, and the expression of pBD2 in Escherichia coli is low, probably because a different gene codon is used by prokaryotic organisms to that used by eukaryotes. Codon preference optimization is one of the ways to increase heterologous expression of pBD2. To achieve high expression of pBD2, the pBD2 gene was redesigned according to the preferred codon in E. coli without altering the amino acid sequence. The optimized gene was inserted into expression vector pET-30a and transformed into E. coli BL21 (DE3) plysS. Our results showed that pBD2 was expressed as His-Tag fusion protein at a level that was approximately 4-6 times greater than from the native gene, based on total protein expression. Expressed fusion pBD2 showed antimicrobial activity against both E. coli and Staphylococcus aureus. Moreover, pBD2 showed weak hemolytic activity and strong heat resistance. These results indicate that fusion pBD2 is functional and has similar properties to those of pBD2 from the native gene. Our current study demonstrated that codon optimization could enhance pBD2 expression in E. coli without altering its function. Therefore, the expression of pBD2 after codon optimization in heterologous host cells might be useful and is worthy of further research.

Clinical observation of umbilical cord mesenchymal stem cell treatment of severe systolic heart failure.

The aim of this study was to evaluate the effectiveness of umbilical cord mesenchymal stem cells (MSCs) in the treatment of chronic systolic heart failure. Fifty-nine hospitalized patients with heart failure were randomly divided into a treatment group (30 patients) and a control group (29 patients). The treatment group received treatment with medication as well as intracoronary transplantation of umbilical cord MSCs, and the control group, only medication. The cardiac structure, function change, and rehospitalization and mortality rates of the 2 groups were observed before and 1 and 6 months after treatment. One month after the transplantation of umbilical cord MSCs, the incidence of fatigue, chest tightness, and dyspnea was high in the treatment group. The 6-min walking distance of the treatment group was found to be significantly higher than that of the control group (P < 0.05); in addition, the NT-proBNP level, left ventricular ejection fraction, and mortality rate of the treatment group were statistically lower than those of the control group (P < 0.05). Readmission rates showed a downward trend, but the difference was not statistically significant (P > 0.05). Using umbilical cord MSCs in the treatment of congestive heart failure can help improve cardiac remodeling and cardiac function and reduce the mortality rate.

Preparation of polyclonal antibody against porcine beta defensin 2 and identification of its distribution in tissues of pig.

Porcine ß-defensin 2 (pBD2) is an antimicrobial peptide in pigs that plays an important role in the immune system by preventing bacterial invasion. To produce an anti-pBD2 antibody, which is not commercially available, we expressed and purified a soluble, his-tagged version of pBD2 (his-pBD2). Purified pBD2 was injected into New Zealand white rabbits to generate polyclonal antiserum. Anti-pBD2 antibodies were purified by ammonium sulfate precipitation, followed by diethylaminoethyl cellulose ion-exchange chromatography. The purified polyclonal antibody showed high sensitivity, with a titer as high as 204,800 by enzyme-linked immunosorbent assay, and it also showed high specificity for both his-pBD2 and native pBD2, as assessed by western blotting. Furthermore, immunohistochemistry analysis using the purified antibody revealed that pBD2 protein is distributed in the tongue, liver, kidney, small intestine, and large intestine of pigs. These results indicate that the prepared polyclonal antibody will be a useful tool for further studies of the function and mechanism of pBD2.

Adaptive protection against damage of preconditioning human umbilical cord-derived mesenchymal stem cells with hydrogen peroxide.

Adaptive protection against damage to human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) was investigated by preconditioning with low-concentration hydrogen peroxide (H2O2) for a short-time period. Separation, culture, amplification, purification, and identification of immunophenotype and growth curve measurements of hUC-MSCs were performed in vitro. At the logarithmic phase, hUC-MSCs were incubated with different (H2O2) concentrations for 1 and 12 h, and the effects were detected by a cell metabolism assay. Then, hUC-MSCs were preconditioned with 10, 20, 50, and 100 mM (H2O2) for 1 h, restored for 0, 12, and 24 h, and then damaged with 700, 800, and 900 mM (H2O2) for 12 h. Cell morphology, cell metabolism, and the number of cells were measured to determine the protective role of preconditioning. Flow cytometry analysis revealed that the cells expressed CD29 and CD44, but not CD34 and CD45. The growth curve showed that hUC-MSCs reached the logarithmic phase in 3-6 days. The cell metabolism assay showed that (H2O2) induced hUC-MSCs damage in a dose- and time-dependent manner. The cell morphology, cell metabolism, and number of cells all showed that preconditioning with 10, 20, 50, and 100 mM (H2O2) for 1 h and restoration for 12 h prevented subsequent damage with 700, 800, and 900 mM (H2O2) on hUC-MSCs. Preconditioning with low-concentration (H2O2) for a short time has a protective effect of preventing damage on hUC-MSCs exposed to high-concentration (H2O2) for a long time, which is dependent on H2O2 concentration and the time interval between preconditioning and damage.