Objective To investigate the effect of hypertonic saline on cerebral water content, tumor necrosis factor-α (TNF-α) level and neuronal apoptosis following focal cerebral ischemia-reperfusion (IR) injury in rats and explore the mechanisms involved. Methods Ninety-six rats were randomized equally into 4 groups, namely the shame-operated group, untreated IR injury group, and 4.2% and 7.5% hypertonic saline groups (HS-A and HS-B groups, respectively). In the latter 3 groups, cerebral ischcmia was induced by middle cerebral artery occlusion for 2 h followed by administration of the corresponding treatments. Serum sodium concentration was measured at 5 min before and at 30, 60 and 90 min after the reperfilsion. At 22 h of rcperfusion, the rats were sacrificed after neurological deficit evaluation, and brain edema was assessed by measuring the wet-to-dry weight ratio of the brain tissue. TNF-α expression in the ischemic brain tissue was measured by enzyme-linked immunosorbent assay (ELISA), and the neuronal apoptosis was analyzed using TUNEL assay. Results In the saline-treated rats, serum sodium level increased significantly after saline administration, lasting for 60 min before recovering the normal levels in HS-A group and for over 90 min in HS-B group. Compared with that in the sham-operated group, the brain water content in rats of the IR group increased in both of the hemispheres, but more obviously in the ischemic hemisphere. In the two saline-treated groups, the water content decreased significantly in the bilateral hemispheres, which was especially obvious in the ischemic hemisphere;administration of 7.5% saline resulted in greater water content reduction in the ischemic hemisphere than 4.2% saline. Compared with the IR group, the two saline-treated groups showed significant reduction in TNF-α levels and apoptotic cells in the brain along with decreased neurological deficits. Conclusion Hypertonic saline can ameliorate cerebral focal IR injury by decreasing the cerebral water content, TNF-α level and neuronal apoptosis following the injury.

OBJECTIVETo obtain the monoclonal antibody against hexon protein of human adenovirus.

METHODSBALB/c mice were immunized with purified recombinant hexon protein, and the spleen cells of the mice were isolated and fused with myloma cells. Four hybridoma cell strains were screened by indirect ELISA and cultured, and the sensitivity, specificity and virus neutralizing activity were analyzed with ELISA, Western blotting and neutralizing test.

RESULTSThe mouse ascites produced by these hybridoma cells contained specific monoclonal antibodies against hexon protein of human adenovirus as identified by ELISA and Western blot, and the antibody generated by 4C6 strain showed human adenovirus type 3-neutralizing activity.

CONCLUSIONThe monoclonal antibodies against hexon protein with high specificity have been successfully obtained, and these antibodies can be useful in developing assays for early diagnosis of HAdV3 infection and also in study of therapeutic drugs of the infection.

Adenoviruses, Human ; chemistry ; immunology ; Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Viral ; biosynthesis ; immunology ; Blotting, Western ; Capsid Proteins ; biosynthesis ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; immunology

The aim of this study was to establish a diagnostic method for ABO genotyping and to investigate the distribution of ABO genotype in Beijing Han population so as to understand the distribution characteristics and regularity of ABO genotype. An ABO genotyping method was established by using multiplex-PCR-RFLP and PCR-SSP techniques, and the ABO allele frequency in Beijing Han population was investigated. The results showed that A102, O1 and B allele were more common genes in Beijing Han individuals. And A102 allele was predominant in the phenotype A group in this population. Three O2 alleles were found and no A201 allele was found while gene frequency investigation was performed. No A101A101, A101O2, A102O2, BO2 and O2O2 in this population were discovered. It is concluded that the primary regularity of ABO allele distribution in Beijing Han population is found through this study. It provides basic reference for further study of ABO types.
ABO Blood-Group System ; genetics ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Young Adult

Limitations of polyacrylamide gel or agarose gel electrophoretic methods in genotyping research affect the interpreting of detection results. In order to develop a simple and reliable method for appraising results of ABO genotyping detection, the microfluidic chip analysis system was established by using microfluidic chip to replace the gel electrophoresis and combining with multiplex-PCR-RFLP technique. 150 blood samples were tested by this microfluidic chip analysis system with multiplex-PCR-RFLP technique to evaluate its stability and accuracy. The results showed that all the testing results were consistent with serologic ABO genotyping results and 1 blood sample with decrease of B antigen caused by CML was identified. In conclusion, the established microfluidic chip analysis system is stable and reliable technique. Application of this technique enables the ABO genotyping results to be more objective and accurate.
ABO Blood-Group System ; genetics ; Blood Grouping and Crossmatching ; methods ; DNA Primers ; genetics ; Genotype ; Humans ; Microfluidic Analytical Techniques ; Microfluidics ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo observe the therapeutic effect and mechanism of Naohuandan (NHD) in treating senile dementia (SD).

METHODSClinical study: Fifty-eight patients with SD, whose diagnosis conforms to the Diagnostic Standard of DSM-IV issued by American Association of Psychiatry, were enrolled and randomly assigned into two groups. The 30 patients in the treated group were treated with NHD, 4 capsules each time, 3 times daily. The 28 patients in the control group were treated with Piracetam, 1.6 g each time, 3 times daily. The therapeutic course for both groups was 3 months. The therapeutic efficacy was estimated and compared by comprehensive scores of memory and cognition, scores of Mini-mental State Examination (MMSE) and Activities of Daily Living (ADL). Experimental study: Rats were divided into the control group, the model group and the high-dosage and low-dosage NHD treated groups. The protective effect of NHD on the per-oxidative damage of hippocampal neurons in beta-amyloid protein induced SD model was observed and the related criteria were determined.

RESULTSClinical study showed that both NHD and Piracetam could improve the clinical symptoms of patients, the two medicines showing insignificant difference in total effective rate. But NHD was better in elevating MMSE score and lowering ADL score in patients than Piracetam (P < 0.05 and P < 0.01). Experimental study showed that (1) 24 and 72 hrs after modeling, the activity of SOD and GSH were lower and the level of MDA higher in the model group than those in the control group (P < 0.05 or P < 0.01). Compared with the model group at the corresponding time points, in the high-dosage NHD group, SOD and GSH were higher, MDA was lower (P < 0.05 or P < 0.01); but in the low-dosage NHD group, SOD at the 72nd hr was higher (P < 0.05) and MDA at 24th and 72nd hrs was lower (P < 0.01). And most of the criteria in the high-dosage NHD group was improved better than that in the low-dosage NHD group. (2) The survival rates of neurons in various groups were not different significantly (P > 0.05) 24 hrs after modeling, but that in the high-dosage NHD group was significantly higher than that in the model group (P < 0.01) and in the low-dosage NHD group 72 hrs after modeling (P < 0.05).

CONCLUSIONNHD is an effective Chinese herbal preparation for treatment of SD, and its mechanism is related with its inhibition on peroxidative injury and protection on neurons.

Aged ; Aged, 80 and over ; Alzheimer Disease ; drug therapy ; metabolism ; Animals ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Glutathione ; metabolism ; Hippocampus ; cytology ; Humans ; Male ; Malondialdehyde ; metabolism ; Middle Aged ; Neurons ; cytology ; drug effects ; metabolism ; Neuropsychological Tests ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the relationship between the distribution of mononucleotide polymorphism of five regulation regions of alpha-IFN among HLA-DRB1*11 gene episodes and the therapeutic effects of alpha-IFN treatment in chronic hepatitis B patients.

METHODSOne hundred seven chronic hepatitis patients from Nanjing Second Hospital who were treated by alpha-IFN for 12 months and then followed at least six months without the treatment were randomly selected for this regressive analysis. They were grouped into a continuous responsive group and a non-continuous responsive group. Hepatitis B virus X interacting protein gene locus was searched in NCBI. Single nucleotide polymorphism (SNP) gene locus was detected based on a pooling sequencing method. Primer and TaqMan-MGB probes referring to different mononucleotide loci were designed respectively to detect SNP in five regulation regions of alpha-IFN. Then gene sequencing differences between the two groups were analyzed.

RESULTSAmong the 107 cases there were 30 cases (28.0%) in the continuous responsive group and 77 cases (71.9%) in the non-continuous responsive group. CT occupation rate in five regulation regions of IFN reached 18.0% in the continuous responsive group and 23.8% in the non-continuous responsive group. AG occupation rate reached 10.8% in the former group and 15.8% in the latter group. The differences in CT and AG between the two groups were significant.

CONCLUSIONSThe distribution of mononucleotide polymorphism of five regulation regions of alpha-IFN among HLA-DRB1*11 gene episodes affects the IFN anti-virus treatment. Detecting the gene distribution of mononucleotide in five regulation regions of alpha-IFN helps in predicting the therapeutic effects of alpha-IFN.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; DNA, Viral ; Genotype ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; genetics ; Humans ; Interferon-alpha ; therapeutic use ; Polymorphism, Single Nucleotide ; Regression Analysis ; Treatment Outcome ; Young Adult

Objective To compare differences of clinical factors related to early pregnancy loss between invitro fertilization-embryo transfer (IVF-ET) treatment and natural pegnancy. Methods A retrospective analysis was performed on the 363 cases of early pregnancy loss between Dec. 2015 to May 2016 in Peking University Third Hospital, during which 173 cases were after IVF-ET treatment(IVF-ET group), and others were natural pregnancies(natural group). Results The average age in IVF-ET group was significantly higher than that in the natural group [(34.1±4.3)versus(31.8±4.1)years old, P<0.01]. The terminating time of pregnancy loss in IVF-ET group was short than that in the natural group [(59.8±9.2) versus(69.9 ± 11.1)days, P<0.01]. The incidence of embryo abnormal chromosome in IVF-ET group was significantly lower than that in the natural group [57.2%(99/173)versus 74.2%(141/190), P<0.01], during which abnormal chromosome numbers were the most common. Conclusions The pregnancy loss of early pregnancy is mainly caused by chromosome abnormality. The proportion of chromosome abnormality in early pregnancy loss after IVF-ET is not higher than that of natural pregnancy, indicating that there are relatively reliable gametes and embryo safety in IVF treatment.

Superoxide Dismutase (SOD)(EC 1.15.1.1)is a metalloenzyme that is found in almost all organisms and catalyzes the dismutation of superoxide anion radical to hydrogen peroxide and molecular oxygen. Three unique and highly compartmentalized mammalian SOD have been biochemically and molecularly characterized to date: Cu, Zn superoxide dismutase (CuZnSOD, SOD1), MnSOD (Manganese Superoxide Dismutase, SOD2)and EC-SOD (Extracellular Superoxide Dismutase, SOD3). Cu, Zn superoxide dismutase (CuZnSOD, SOD1)is a copper and zinc-containing homodimer that is found almost exclusively in intracellular cytoplasmic spaces. CuZnSOD is widely distributed and comprises about 90% of the total SOD. Cytoplasmic and periplasmic SOD exists as dimers,whereas chloroplastic and extracellular enzymes exist as tetramers. Structure supports independent functional evolution in prokaryotes and eukaryotes. CuZnSOD are thought to protect the brain, lungs, and other tissues from oxidative stress. This paper reviewed the gene, molecular and chemical structure and biological function of CuZnSOD.

G-rich oligonucleotides (GROs) belong to a novel class of phosphodiester oligonucleotides. They can form G-tetramer structure which contributes to cell cycle arrest and growth inhibitory effects by non-antisense pathway. This study was aimed to investigate the biological effects of GRO-26B on leukemia cell lines. Cell proliferation of different cell lines were detected by using MTT method and trypan blue incorporation assay. Alteration of cell cycle was analyzed by using flow cytometry. Apoptosis was detected by using Annexin V/PI kit. Western blot was used to detect the expression level of cyclins and CDKs. Morphological features of GRO-26B-treated cells was observed by light microscopy and transmission electron microscopy (TEM). The results showed that GRO-26B could inhibit the proliferation of AML cell lines, such as U937 and NB4 cells in a dose-dependent manner. GRO-26B induced the cell cycle to be arrested at S phase in time-dependent manner, which was associated with the alteration of cyclin A, cyclin B, CDC2 and CDK2. The morphology of cells treated by GRO-26B also showed a distinct change as compared to the untreated cells. It is concluded that GRO-26B can inhibit AML cell proliferation, which is partially associated to cell cycle arrest at S phase. The S phase arrest is related to cyclins/CDKs. The regulation mechanism of cell cycle and proliferation is complicated. All of the above-mentioned phenomena need to be studied in the future.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Leukemia ; pathology ; Oligonucleotides ; pharmacology