Objective:To investigate the effect of staphylococcus aureus on expressions of inducible nitric oxide synthase (iNOS) and interleukin-1 beta (IL-1β) in THP-1 monocytes under high glucose concentrations. Methods:THP- 1 monocytes were incubated at different surroundings with 2×2 factorial design. There were 4 experimental groups in the study, which were analyzed by univariate analysis of variance. The total RNA was distilled. The expressions of iNOS and IL-1β were examined by SQ-RT-PCR. Results:The expressions of iNOS and IL-1β were affected by high glucose concentration, bacteria and both of them together in THP-1 monocytes. The expressions of iNOS and IL-1β were weakened in high glucose concentration compared with that of control(P < 0.01). The expressions of iNOS and IL-1β were weakened in the high glucose concentration and bacterial infection compared with that of the high glucose concentration only(P < 0.01). The expressions of iNOS and IL-1β increased in bacterial infection compared with that of control(P < 0.01). The expression of IL-1β increased in high glucose concentration and bacterial infection compared with that of the high glucose concentration only(P < 0.01) but the expression of iNOS was not distinct. Conclusion:When staphylococcus aureus infected monocytes, the expressions of iNOS and IL-1β were weakened in high glucose concentration, which suggested that the depressed function of immunocyte was related with high glucose concentration.

Objective To study the effects of granisetron hydrochloride on vasovagal syncope(VVS) in rabbits.Methods Twenty-four healthy New Zealand rabbits were divided stochastically into control group and intervention group,12 in each group. The control group was injected intravenously with normal saline. The intervention group was injected intravenously with granisetron hydrchloiride.Rabbit VVS models were established,each was taken at 4 points in time in the bloodletting process:T1,T2,T3,T4,to compare the bloodletting time,the concentration of 5-hydroxytryptamine(5-HT) in T2,T3,T4 and the total blood volume between the groups,and monitor the heart rate, blood pressure during the entire process.Results 1.The time of intervention group in T2,T3,T4 was longer than the time of control group obviously(P

To explore the epidemiological character of Methicillin-resistant Stapkylococcus aureus (MRSA) by the phenotyping and genotyping motheds and to investgate the source, transmission, and the spread of nosocomial MRSA infection, consequently, reducing the nosocomial infection of MRSA. In this study, 19 MRSA strains were isolated from patients and environment in a hospital in two months. Patterns of resistantce against 16 antimicrobial agents and pulsed-field gel electrophoresis ( PFGE) of these strains were analyzed to find the relationship among those isolates Clustering analysis was made from the patterns. Some isolates with high homology was found in 19 MRSA, 11 of them belong to type A, and 8 of them belong to the same subtype A1. They were endemic in burn ward, oncological ward and ICU. In addition, 4 isolates were clustered into group B, all found in the same ward of burn unit Thus, our results indicated a outbreak of MRSA ( A type) in this hospital and the potential prevalence of MRSA (B type) , which might be mediated by health care stuff. It is essential to enhance the infection control implementation and to utilize the PFGE genotyping system for the real-time surveillance of MRSA.

We used metabolomics to investigate the ability of a traditional Mongolian medicine called modified Tabusen-2 (MT-2) to improve kidney yang deficiency (KYD) in rats. All animal experiments were conducted under the guidance and standards of the Medical Ethics Committee of Inner Mongolia Medical University. SD rats were divided into 6 groups of six rats: a normal group, a model group, Jinkuishenqi pill administration group (1.26 g·kg^-1), and MT-2 administration in high-, medium- and low-dose groups (1.512, 0.756, and 0.378 g·kg^-1). KYD was established by intramuscular injection of hydrocortisone (HC) and biochemical indicators and clinical characterization was used to confirm that KYD was established. All groups received intragastrically administered drug (Jinkuishenqi pill or MT-2) or saline. Serum from each group was collected after 8 weeks and analyzed by UPLC-Q-exactive-MS to measure various biochemical indicators. The biomarkers affected by MT-2 were identified and the metabolic pathways of KYD regulated by MT-2 were analyzed by metabolomic analysis. The results show that MT-2 can decrease serum creatinine (Cr) in KYD rats and significantly increase (P<0.05) the content of thyroid stimulating hormone (TSH) and luteinizing hormone (LH). In serum samples, 38 biomarkers such as corticosterone, L-phenylalanine, and DL-tryptophan were measured as possible indicators for disease development in KYD rats. MT-2 lowered 18 biomarkers of KYD, including corticosterone, deoxycorticosterone, and L-phenylalanine, and altered 13 related metabolic pathways including phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and steroid hormone biosynthesis, resulting in an overall improvement in KYD. MT-2 appears to be important in improving KYD in rats mainly by regulating metabolites such as amino acids, steroids and lipids.

Adult ; Female ; Humans ; Liver ; immunology ; pathology ; Liver Cirrhosis, Biliary ; immunology ; pathology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology

Objective To study the apoptosis of facial motor neurons and the expression of apoptosis-related genes, Bcl-2 and Bax, in the animal model of viral facial paralysis. Methods Total of 84Balb/c mice were divided into viral inoculation group and nerve transaction group. The animals were executed 1, 3, 7, 10, 15, 20 and 30 days after being operated respectively. The histopathological features of facial neurons in brain stem were observed by HE and Nissl stain. The changes of facial neuronal apoptosis were observed by TUNEL The changes of expression of Bcl-2 and Bax genes in facial neurons were observed by immunohistochemistry staining. Results After nerve transection, increased apoptotic cells were found in homolateral facial motor neucleus and the peak appeared at 10 and 15 days. The level of Bcl-2 expression inneurons declined while the expression of Bax increased gradually. Correspondingly, the ratio of Bcl-2/Bax declined. In the viral inoculation group, no visible change of apoptosis and Bax expression, but the level of Bcl-2 and the ratio of Bcl-2/Bax increased gradually. Conclusions Comparing to axotomy, facial motor neucleus in HSV-1 infective animal model are free of apoptosis. Both the mild form of lesion and the ability to block apoptosis of HSV-1 are likely to be involved into the phenomenon. Bcl-2 and Bax might interfere with the apoptotic response.

Objective To investigate the expression and distribution of intrahepatic CD4+ CD25+regulatory T cells in immuno-tolerant and immuno-elearanee phase of patients with chronic hepatitis B. Methods The expression of FoxP3 was detected in 19 cases of immuno-tolerant phase and 12 cases of immuno-clearance phase by immunohistochemistry. The relation between the intrahepatic expression of FoxP3 and the clinico-pathological features were analyzed. Results The positive signal of FoxP3 is located in nuclear of lymphocyte and mainly aggregated in portal areas as well as occasionally scattered in hepatic sinusoids. The expression of intrahepatic FoxP3 in the group of immuno-tolerant phase was significantly increased than those in normal control(P<0.01), and greatly decreased than those in immuno-clearance phase (P<0.01). No correlation was observed among the expression of intrahepatic FoxP3, ALT, levels of HBV DNA, HBeAg positive, in patients of immuno-clearanee phase, respectively. There were significant differences between immuuo-tolerant phase and immuno-clearance phase age, ALT, TBIL, PTA, HBV-DNA and detection of HBeAg but not in sex and family history of HBV infection. Conclusion CD4+ CD25+ regulatory T cells may play important roles in the clearance of HBV as well as in liver inflammation and injury during chronic HBV infection.

BACKGROUNDCytosine deaminase (CD) converts 5-fluorocytosine (5-FC) to 5-fluorouracil (5-FU) in CD/5-FC gene therapy, 5-FU will be mostly converted into nontoxic beta-alanine without uracil phosphoribosyltransferase (UPRT). UPRT catalyzes the conversion of 5-FU to 5-fluorouridine monophosphate, which directly kills CD::UPRT-expressing cells and surrounding cells via the bystander effect. But the pharmacokinetics and the bystander effect of CD::UPRT/5-FC has not been verified in vivo and in vitro. Before the CD::UPRT/5-FC bi-gene therapy system is used in clinical trial, it is essential to monitor the transgene expression and function in vivo. Thus, we developed a preclinical tumor model to investigate the feasibility of using (19)F-magnetic resonance spectroscopy ((19)F-MRS) and optical imaging to measure non-invasive CD and UPRT expression and its bystander effect.

METHODSC6 and C6-CD::UPRT cells were cultured with 5-FC. The medium, cells and their mixture were analyzed by (19)F-MRS. Rats with intracranial xenografted encephalic C6-CD::UPRT glioma were injected intraperitoneally with 5-FC and their (19)F-MRS spectra recorded. Then the pharmacokinetics of 5-FC was proved. Mixtures of C6 and C6-CD::UPRT cells at different ratios were cultured with 5-FC and the cytotoxic efficacy and survival rate of cells recorded. To determine the mechanism of the bystander effect, the culture media from cell comprising 25% and 75% C6-CD::UPRT cells were examined by (19)F-MRS. A comparative study of mean was performed using analysis of variance (ANOVA).

RESULTS(19)F-MRS on samples from C6-CD::UPRT cells cultured with 5-FC showed three broad resonance signals corresponding to 5-FC, 5-FU and fluorinated nucleotides (F-Nuctd). For the C6 mixture, only the 5-FC peak was detected. In vivo serial (19)F-MRS spectra showed a strong 5-FC peak and a weak 5-FU peak at 20 minutes after 5-FC injection. The 5-FU concentration reached a maximum at about 50 minutes. The F-Nuctd signal appeared after about 1 hour, reached a maximum at around 160 minutes, and was detectable for several hours. At a 10% ratio of C6-CD::UPRT cells, the survival rate was (79.55 +/- 0.88)% (P < 0.01). As the C6-CD::UPRT ratio increased, the survival rate of the cells decreased. (19)F-MRS showed that the signals for 5-FU and F-Nuctd in the culture medium increased as the ratio of C6-CD::UPRT in the mixture increased.

CONCLUSIONS(19)F-MRS studies indicated that C6-CD::UPRT cells could effectively express CD and UPRT enzymes. The CD::UPRT/5-FC system showed an obvious bystander effect. This study demonstrated that CD::UPRT/5-FC gene therapy is suitable for 5-FC to F-Nuctd metabolism; and (19)F-MRS can monitor transferred CD::UPRT gene expression and catalysis of substrates noninvasively, dynamically and quantitatively.

Animals ; Antimetabolites ; pharmacokinetics ; therapeutic use ; Cell Line ; Cytosine Deaminase ; genetics ; physiology ; Flucytosine ; pharmacokinetics ; therapeutic use ; Genetic Therapy ; methods ; Glioma ; drug therapy ; therapy ; Humans ; Magnetic Resonance Imaging ; Male ; Pentosyltransferases ; genetics ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo detect the methylation status of 5'CpG island in the core promotor of maspin gene in RKO human colorectal cell line,and to explore the transcription regulation of DNA 5'CpG island demethylation on maspin tumor suppressor gene and its effect on the growth of cancer cell.

METHODSThe status of 5 'CpG island methylation of maspin gene in RKO human colorectal cell line was analyzed using methylation specific polymerase chain reaction (MSP). After treated with a specific demethylating agent, 5-Aza-2'-deoxycytidine, reverse transcription polymerase chain reaction (RT- PCR) was used to examine maspin gene expression. Cell proliferation was evaluated using MTT assay,distribution of cell cycle and rate of apoptosis were determined using flow cytometry.

RESULTSThe 5'CpG island methylation in the core promotor of maspin gene was detected in RKO human colorectal cell line. After treatment with three different concentration of 5-aza-2'-deoxycytidine, the expression of maspin mRNA increased 10.89, 16.91, 23.97 times respectively. MTT array showed the proliferation activity of RKO cell line was obviously reduced after 5-aza-2'-deoxycytidine treatment. The cells were arrested in G(0)/G(1) phase,and the apoptosis rates were 5.17%, 8.71% and 11.23% respectively compared with control group.

CONCLUSIONThe 5'CpG island methylation is probably responsible for maspin expression silencing in RKO human colorectal cell line, 5-aza-2'-deoxycytidine may effectively cause demethylation and inhibit the growth of tumor cell by reactivating the gene transcription silenced by aberrant hypermethylation.

Azacitidine ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; CpG Islands ; drug effects ; DNA Methylation ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Serpins ; genetics ; Transcription, Genetic ; drug effects

OBJECTIVETo investigate the association among serous and cerebrospinal fluid (CSF) TNF-alpha level, gene polymorphisms of TNF-alpha and multiple sclerosis (MS) in Han nationality of southern China.

METHODSMS diagnosis was base on Poser (1983) criteria. Fifty-five patients with nonimmulogical diseases and 68 patients with MS from southern China were enrolled in the study, and their TNF-alpha level of serum and CSF were measured by double antibody sandwich ABC-ELISA. TNF-alpha -308G/A in 106 normal healthy subjects and 68 MS patients was genotyped with polymerase chain reaction-restriction fragment length polymorphism.

RESULTSThere was significant difference in the serous TNF-alpha level between nonimmune patients and active MS patients (234+/- 76 pg/mL vs 276+/- 71 pg/mL, P< 0.05), but not in the CSF (245+/- 83 pg/mL vs 265+/- 78 pg/mL, P> 0.05). The gene frequency distribution of TNF-alpha -308G/A was corresponding with Hardy-Weinberg equilibrium. The positive rate of genotype AA and the gene frequency of allele A of TNF-alpha were 4.4% and 14.0% in MS group, and 0 and 8.50% in healthy subjects, there was no statistical significance (P> 0.05).

CONCLUSIONThe TNF-alpha level in serum is associated with active MS, but not in the CSF. The gene polymorphisms of TNF-alpha -308G/A is not associated with MS in Han nationality of southern China.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; China ; Enzyme-Linked Immunosorbent Assay ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Multiple Sclerosis ; blood ; cerebrospinal fluid ; genetics ; Polymorphism, Genetic ; Tumor Necrosis Factor-alpha ; blood ; cerebrospinal fluid ; genetics