Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Brain ; drug effects ; pathology ; Disease Models, Animal ; Estrogens ; pharmacology ; Female ; Hypoxia-Ischemia, Brain ; drug therapy ; In Situ Nick-End Labeling ; Male ; Rats ; Rats, Wistar

Aim To study the effect of the total flavonoids of turpinia arguta seen (TFS) on immune function of rats with adjuvant arthritis. Methods Adjuvant arthritis (AA) was induced on d0 by intradermal injection of Complete Freund′s Adjuvant (FCA), containing 10 mg heat-inactive BCG in 1ml paraffin oil. On d 28 after immunization, AA rats were killed by cervical decollation. Immune cells (splenocytes and peritoneal macrophages) were obtained, then cultured in vitro with TFS. Some immune indexes were detected respectively such as the splenocytes proliferation and the productions of IL-1 and IL-2 were measured by the method of cell proliferation, the production of prostaglandin E2 was determined by radioimmunoassay. Results Compared with normal groups, the response of splenocytes to ConA and LPS was lowered, IL-2 syn-thesis was decreased, and IL-1 and PGE_2 released from PM? were elevated. TFS (10~ -8 ~10~ -4 mg?L~ -1 ) not only increased splenocytes proliferation, which induced by ConA and LPS, and IL-2 production of splenocytes, but also reduced the elevated productions of IL-1 and PGE_2 released from peritoned macrophate in AA rats in vitro. Conculsion TFS could adjust abnormal immune function in AA rats.

The present study was designed to observe the influence of cerebral ischemia/reperfusion injury on learning and memory in hyperlipidemic rats and estimate the changes of activity of autonomic nervous system. Twenty-three male Wistar rats were randomly divided into three groups, named control group (C group, n=10), hyperlipidemia group (H group, n=6) and hyperlipidemia-ischemia group (HI group, n=7), respectively. The rats in H and HI group were fed a high-fat diet for 2 weeks and the rats in all groups were examined through Morris water maze (MWM) task. The rats in HI group underwent ischemia/reperfusion by 2-vessel occlusion (2-VO) method, and had electrocardiogram (ECG) recording simultaneously. The MWM task and ECG recording were taken again after 7 d of recuperation. The following results were obtained: (1) In the second place navigation performance and probe trial performance, the frequency of memory in quadrant of hidden-platform and memory score decreased significantly in HI group compared to that in C and H groups. (2) The heart rate in HI group decreased slowly after ischemia; the power at high frequency band (HF) reduced gradually, meanwhile the power at middle frequency band (MF) and the ratio of power at MF and HF decreased clearly compared to baseline value. (3) After 7 d of ischemia/reperfusion, the heart rate in HI group was significantly higher than that in H group (P<0.05). While there was no statistical change in the power at MF, the power at HF decreased and the ratio of MF/HF increased significantly (P<0.05). The data demonstrated that ischemia/reperfusion decreased the activity of autonomic nervous system, and the reduction of sympathetic nerve activity was much more than that of vagus nerve activity. The results suggest that the hippocampus neuron injury caused by ischemia induces cognitive disorder and imbalance of vago-sympathetic nerve activity accompanied by vagus nerve suppression.
Animals ; Autonomic Nervous System ; physiopathology ; Brain Ischemia ; etiology ; physiopathology ; Heart Rate ; physiology ; Hippocampus ; physiopathology ; Hyperlipidemias ; complications ; Learning Disorders ; etiology ; physiopathology ; Male ; Memory Disorders ; etiology ; physiopathology ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; pathology ; physiopathology

OBJECTIVETo investigate the expression of HER2/neu, Ki-67 and TK1 protein in meningiomas in correlation with tumor grades and recurrence.

METHODSTwenty cases of each of the following types of meningiomas were selected for the study, namely: the benign non-recurrent, recurrent benign, atypical and malignant, according to the World Health Organization (WHO) histological classification of nervous system, 2007. Immunohistochemistry study for HER2/neu, Ki-67 and TK1 protein was performed. HER2/neu gene amplification was detected using FISH. Cases with HER2 protein overexpression were studied by immunohistochemistry staining. The results of the biomarker assays were also used to study the correlationship with the tumor grades and tumor recurrency.

RESULTSImmunohistochemistry showed that the positive rates of HER2 expression in non-recurrence benign group, recurrence benign group, atypical group and malignant group were 3 cases (15%), 6 cases (30%), 7 cases (35%), and 10 cases (50%), respectively (P < 0.05). A higher tumor grade was correlated with a higher rate of HER2/neu expression. The Ki-67 and TK1 labeling index (LI) in non-recurrence group were lower than those in the atypical or malignant group (P < 0.05), whereas the atypical group had lower LI than that of the malignant group (P < 0.05). Higher levels of LI of Ki-67 and TK1 were correlated with higher tumor grades and recurrence of the benign meningiomas (P < 0.05). Expression of HER2 was positively correlated with Ki-67 and TK1 (r = 0.445, P < 0.01; r = 0.501, P < 0.01, respectively), and there was a positive correlation between Ki-67 and TK1 (r = 0.450, P < 0.01) as well. HER2/neu gene copy amplification in 7 of 26 cases (26.9%) of HER2 immunopositive meningiomas. The rates of HER2/neu gene amplification were 0 in tumors with 1+ immunopositivity, 4/6 in tumor with 2+ immunopositivity and 3/4 in tumor with 3+ immunopositivity. HER2/neu gene amplification in 3+ and 2+ immunopositive cases had no statistical significance (P > 0.05). Aneuploidy of chromosome 17 existed in 9 of 26 of HER2 immunopositive meningiomas (34.6%). However, the rates of chromosome 17 aneuploidy had no significant difference among tumors with variable HER2/neu imumopositivity (P > 0.05).

CONCLUSIONSHigh levels of HER2 and Ki-67 or TK1 expression correlate with the increase of tumor grades and tumor recurrence. HER2/neu gene amplification is seen in a subset of meningiomas with the protein expression (26.9%). A combination of biomarker study including HER2/neu, Ki-67 and TK1 may be useful in predicting the biological behavior of meningiomas.

Aneuploidy ; Chromosomes, Human, Pair 17 ; Female ; Gene Amplification ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Ki-67 Antigen ; metabolism ; Male ; Meningeal Neoplasms ; genetics ; metabolism ; pathology ; Meningioma ; genetics ; metabolism ; pathology ; Middle Aged ; Neoplasm Recurrence, Local ; Receptor, ErbB-2 ; genetics ; metabolism ; Thymidine Kinase ; metabolism

OBJECTIVE: To construct a technological platform of 2-dimensional tumor microvascular architecture phenotype (2D-TAMP) expression. METHODS: Thirty samples of non-small cell lung cancer (NSCLC) were collected after surgery. The corresponding sections of tumor tissue specimens to the slice of CT perfusion imaging were selected. Immunohistochemical staining,Gomori methenamine silver stain, and electron microscope observation were performed to build a technological platform of 2D-TMAP expression by detecting the morphology and the integrity of basement membrane of microvasculature, microvascular density, various microvascular subtype, the degree of the maturity and lumenization of microvasculature, and the characteristics of immunogenetics of microvasculature. RESULTS: The technological platform of 2D-TMAP expression was constructed successfully. There was heterogeneity in 2D-TMAP expression of non-small cell lung cancer. The microvascular of NSCLC had certain characteristics. CONCLUSION 2D-TMAP is a key technology that can be used to observe the overall state of micro-environment in tumor growth.
Capillaries ; ultrastructure ; Carcinoma, Non-Small-Cell Lung ; blood supply ; pathology ; Humans ; Lung Neoplasms ; blood supply ; pathology ; Neovascularization, Pathologic ; pathology ; Phenotype ; Regional Blood Flow ; physiology

OBJECTIVEReal-time RT-PCR, cell culture and embryonated eggs culture for influenza detection were compared by analyzing the data of influenza surveillance in Shenzhen in second half of 2009.

METHODS1092 clinical samples (throat swabs) collected during second half of 2009 were tested by real-time RT-PCR, cell culture and embryonated eggs culture, and the results were analyzed by statistical methods.

RESULTSThe positive rate were 54.21%, 27.11% and 16.21% using real-time RT-PCR, cell culture and embryonated eggs culture, and the sensitive were 100%, 50% and 29.9%. The lowest dilutions of virus detected by real-time RT-PCR were 10(-2) TCID50/ml.

CONCLUSIONThe sensitive of real-time RT-PCR was higher than culture and the specificity was also very high. It was more suitable for emergency detect. The sensitive of cell culture for H3N2 subtype was higher, and sensitive of embryonated eggs culture for type B was higher.

Animals ; Chick Embryo ; Humans ; Orthomyxoviridae ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Virus Cultivation ; methods

Adolescent ; Child ; China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Influenza A virus ; isolation & purification ; Influenza, Human ; epidemiology ; Male ; Prevalence

The use of tumor antigen specific antibody for the delivery of therapeutic agents offers the possibility of targeting therapy with reduced toxicity to normal tissues compared to conventional treatments. In previous work, the human-mouse chimeric antibody fragment Fab' directed against CD20 was constructed from the new anti-CD20 antibody HI47 (a mouse IgG3, K). The chimeric antibody fragment Fab' could reduce its antigenicity, but the yield, quality and affinity of chimeric antibody fragment Fab' restrict its use. To improve affinity of chimeric antibody fragment Fab', a new phasmid pYZcpp3, which expresses chimeric antibody fragment F(ab')2, was constructed by adding a sequence encoding a small peptide, (CPP)3, to C-terminus of heavy chain constant region of chimeric antibody fragment Fab'. Using the pYZcpp3 to transform E. coli. 16c9, the genetically engineered bacteria 10916# was obtained. 10916# can secret the soluble chimeric antibody fragment Fab' and F(ab')2 into periplasmic. The yield was up to 360 mg/L with the percent of F(ab')2 up to 45% in 19L fermentor by the high density fermentation technology. Without denaturation and renaturation, the F(ab')2 has possessed the native three-dimensional structure. The purity of F(ab')2 was more than 90% after the purification of protein G affinity chromatography and S200 size exclusion chromatography. The F(ab')2 could distinguish and bind to Raji cells (CD20+) by FACS. F(ab')2 could inhibit the proliferation of Raji cells in vitro by MTT, IC50 was 22.8 microg/mL. HI47 and its chimeric fragments F(ab')2 induced a significant level of apoptosis (23.5%, 20.8%, respectively), independent of any cross-linking agents, in Raji cells after 24 h incubation. The chimeric antibody fragment F(ab')2 directed against CD20 is possible to apply to tumor therapy in clinic in the future.
Antigens, CD20 ; immunology ; Apoptosis ; Escherichia coli ; genetics ; Fermentation ; Humans ; Immunoglobulin Fab Fragments ; chemistry ; genetics ; isolation & purification ; therapeutic use ; Lymphoma, B-Cell ; therapy ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification ; therapeutic use

OBJECTIVETo investigate the effects of hepatitis B virus (HBV) in semen on human semen parameters and sperm DNA integrity.

METHODSWe detected HBV DNA in the semen samples of 153 HBsAg-seropositive patients by real-time fluorescence quantitative PCR and calculated the sperm nuclear DNA fragmentation index (DFI) by sperm chromatin dispersion (SCD) assay. We compared the semen parameters between the HBV DNA-positive group (A, n = 43) and HBV DNA-negative group (B, n = 110) and analyzed the correlation of sperm DFI with the number of HBV DNA copies in the semen.

RESULTSHBV DNA was detected in 43 (28.1%) of the 153 semen samples. No statistically significant differences were observed in age, semen volume and sperm concentration between groups A and B (P >0.05). Compared with group B, group A showed significantly decreased sperm viability ([58.0 +/- 18.8]% vs [51.4 +/-17.1]%, P<0.05), progressively motile sperm ([29.6 +/- 13.3]% vs [24.5 +/- 10.1]%, P<0.05), average straight-line velocity ([23.7 +/- 4.0] microm/s vs [19.9 +/- 4.5 ] microm/s, P<0.01) and average path velocity ([26.5 +/- 7.0] microm/s vs [23.4 +/- 5.3] microm/s, P<0.01), but remarkably decreased sperm DFI ([19.3 +/- 8.0]% vs [24.2 +/- 9.4]%, P<0.01). The number of HBV DNA copies in semen exhibited a significant positive correlation with sperm DFI (r = 0.819, P < 0.01).

CONCLUSIONHBV DNA in semen is not significantly associated with the number of sperm, but may affect sperm viability, velocity and DFI. There is a load-effect relationship between the number of HBV DNA copies in semen and sperm nuclear DNA integrity.

Adult ; DNA Fragmentation ; DNA, Viral ; isolation & purification ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; Humans ; Male ; Semen Analysis ; Sperm Count ; Spermatozoa ; virology

OBJECTIVETo investigate sperm DNA integrity in male infertility patients with hepatitis B virus (HBV) infection.

METHODSThis study included 90 infertile men with HBV infection (group A), 82 infertile men without HBV infection (group B) and 70 normal fertile men (group C). We detected sperm DNA integrity among the subjects, including DNA fragmentation index (DFI) and high DNA stainability (HDS), by sperm chromatin structure assay (SCSA), and compared them among the three groups.

RESULTSDFI was higher in group A ([28.17 +/- 13.06]%) than in B ([26.64 +/- 9.79]%) and C ([15.67 +/- 4.73]%), significantly higher in A and B than in C (P < 0.05) but with no significant difference between A and B (P > 0.05). HDS was higher in group A ([10.83 +/- 5.601]%) than in B ([9.04 +/- 3.48]%) and C ([8.04-2.25]%), with significant difference between A and C (P < 0.05).

CONCLUSIONSperm DNA integrity of infertile males is significantly different from that of normal fertile men, and infertility with HBV infection further impairs sperm DNA, which is manifested by abnormal sperm nuclear maturity.

Adult ; Case-Control Studies ; Chromatin ; DNA ; genetics ; DNA Damage ; Hepatitis B ; pathology ; Hepatitis B virus ; Humans ; Infertility, Male ; genetics ; virology ; Male ; Sperm Count ; Spermatozoa ; pathology ; Young Adult