Objective To evaluate cerebral autoregulation in cerebral vascular disease using transeranial Doppler(TCD)-CO_2 test and head-upright tilt test(HUTT).Methods Thirty-two patients with occlusive cerebral vascular disease (group A),28 cerebral infarction patients without stenosis (group B), and 26 controls were included.Cerebrovascular reactivity was calculated in terms of the breath-holding index (BHI) and vascular motor reactivity (VMR) by TCD-CO_2 test.Changes of cerebral blood flow velocity (CBFV),blood pressure (BP) and heart rate (HR) were detected during HUTT to assess the cerebral pressure-autoregulation.Results Compared with controls,VMR decreased in group A and B,being 0.18?0.02,0.26?0.04 respectively,and so did BHI,being 0.76?0.15,1.05?0.15 respectively with significance(P

Objective To evaluate the effectiveness of several methods in eliminating Mycoplasma contamination in cell culture of human hepatoma-derived cell line C3A. Methods PCR was performed to detect the Mycoplasmas contamination in cell cultures. The contaminated samples were treated by ciprofloxa-cin, heating, Plasmocure or co-culturing with macrophages. Transmission electron microscope ( TEM) and Q-PCR were used to comparatively analyze the cell morphology and gene expression before and after Plas-mocure treatment. Results Plasmocure succeeded in eliminating Mycoplasma contamination, while cipro-floxacin showed temporary efficacy. Heating and co-culturing with macrophages failed to eliminate Mycoplas-ma contamination. No Mycoplasma contamination in the Plasmocure-treated group was observed under TEM and the expression of ALB, TF and CYP3A4 genes were higher than the genes expressed in the contaminated group (P<0. 01). Conclusion Plasmocure treatment was effective in eliminating Mycoplasma contamina-tion in cell culture. Moreover, the cell morphology and gene expression in Plasmocure-treated group were re-stored to normal.

Improving medical students' calculation ability in statistics has become the focus and difficulty of medical statistics course teaching, and its application relies heavily on statistical calculation software. In order to explore a new teaching approach which combined the advantages of traditional method and web-based calculation, we intended to build a web computing platform applying the Browser/Server (B/S) mode based on the campus network, and to revise the current syllabus of medical statistics, as well as to create a virtual web-lab containing a typical case library. Practice has proved that the new mode effec-tively improved the practice capability of students and changed the traditional teacher-centered teaching approach.

Objective To investigate the correlation between plasma NT-proBNP and the severity of coronary artery disease. Methods One hundred and eighty five subjects were divided into three groups:83 patients in acute coronary syndromes group, 62 patients in stable angina pectoris group and 40 healthy subjects in normal group. The plasma NT-proBNP concentrations were measured by enzyme-linked immunosorbent assay (ELISA) . The plasma NT-proBNP levels of acute coronary syndrome patients and stable angina patients were analyzed. Furthermore, the coronary syndrome patients were divided into three groups according to the result of angiography:single vessel group, dual-vessel group and multiple-vessel group. The relationship of plasma NT-proBNP levels with the severity of coronary artery disease was analyzed. Results (1) In the group of acute coronary syndromes, the plasma NT-proBNP concentrations were obviously higher than the stable angina pectoris group and normal group (P<0.01) . (2) In the groups of coronary heart disease, the plasma NT-proBNP concentrations of the multiple-vesel group were obviously higher than the single vessel group and the dual-vessel group (P<0.01) . The plasma NT-proBNP concentrations of dual-vessel group were obviously higher than the single vessel group (P<0.01) . (3) The plasma NT-proBNP levels and the severity of coronary lesions were positively correlated (rs=-0.796, P<0.01) . Conclusion The plasma NT-proBNP levels is closely related to the severity of coronary artery disease.

AIM:To remark the effect of Qingguang'an Ⅱ on expression of PAX6, Ngn1, and Ngn2 mRNA of rats with chronic high intraocular pressure.METHODS:Totally 40 male SD rats were randomly divided into 6 groups, that was:A:blank group, B:model group, C:Qingguang'an Ⅱ low dose group, D:Qingguang'an Ⅱ moderate dose group, E:Qingguang'an Ⅱ high dose group, F:Yimaikang disket group.B, C, D, E, F groups of experimental rats were established the model of chronic high intraocular pressure (IOP) by cauterizing of superficial scleral vein.Animal model was established successfully by using monitoring IOP consistently keep above 25mmHg for 8wk as cut-off criterion.Tissues of Eyes were obtained after intragastric administration for 2wk and 4wk.The expressions of PAX6, Ngn1, and Ngn2 mRNA were investigated by Real-time PCR.RESULTS:At the time-point of 2wk, PAX6, Ngn1, and Ngn2 mRNA in group B were statistically expressed in lower level comparing with other groups (P<0.05).Moreover, at the time-point of 4wk, PAX6, Ngn1, and Ngn2 mRNA in group C, D and E were statistically expressed in higher level comparing with group F (P<0.05).Besides, PAX6, Ngn1, and Ngn2 mRNA in group C and D were statistically expressed in lower level comparing with group E (P<0.05).PAX6, Ngn1, and Ngn2 mRNA in group C and D were expressed in similar level(P>0.05).CONCLUSION:In summar, Qingguang'an Ⅱ and Yimaikang disket can remarkably increase the expressions of PAX6, Ngn1, and Ngn2, which suggest protecting the optic nerve of rats caused by chronic high IOP.What's more, this study indicated that, in the protection of optic nerve of rats with chronic high IOP, the high dose of Qingguang'an Ⅱ at the time-point of 4wk was the better choice.

The arrenotokous toxicity of triptolide was evaluated, and the rate of sperm abnormality, the changes of the lipid peroxide, the enzyme activity and the hormone in male rats were observed. With the negative and positive control group, the healthy rats were respectively given by gavage triptolide suspension at the dose of 0.025, 0.05, 0.1 mg x kg(-1) for 30 days. Then the rats were killed for the measurement of the indicators in testis and serum, as well as the study on the sperm abnormality. The results showed that the positive control group had significant difference, compared with the negative control group. The content of SOD, LDH, G-6-PD, Na+ -K+ -ATPase, Ca+ -Mg+ -ATPase decreased significantly in 0.05 mg x kg(-1) group, and reduced more obviously with exposure to the dose of 0.1 mg x kg(-1). The levels of GSH-Px and beta-G showed a significant decrease in the testis of rats only at the dose of 0.1 mg x kg(-1). Nevertheless, the MDA levels, the FSH levels and the LH levels showed no significant difference. The deformity rate of sperm increased significantly in 0.05 mg x kg(-1) group and 0.1 mg x kg(-1) group. The results indicated the triptolide had the effect of the lipid peroxidation to damage Spermatogenic cells, Sertolis cells and Leydig cells. At the same time, the triptolide interfered not only with the energy supply process of aerobic and anaerobic glycolysis,but also with the energy utilization in testis by affecting the activities of testis marker enzymes, and produced a damage chain of the male reproductive system
Animals ; Diterpenes ; toxicity ; Drugs, Chinese Herbal ; toxicity ; Epoxy Compounds ; toxicity ; Lipid Peroxidation ; drug effects ; Male ; Organ Size ; drug effects ; Phenanthrenes ; toxicity ; Rats ; Rats, Wistar ; Reproduction ; drug effects ; Spermatozoa ; abnormalities ; drug effects ; metabolism ; Testis ; drug effects ; growth & development ; metabolism ; Tripterygium ; chemistry ; toxicity

In order to obtain active recombinant PNGase F in Escherichia coli, a prokaryotic expression vector pET28a/PNGase F was constructed. Amplification of PNGase F was obtained using PCR technique employing suitable primers designed according to the PNGase F gene sequence from Flavobacterium nmeningosepticum. The expression of PNGase F gene in LB medium or M9 medium led to the formation of inclusion body and soluble protein, respectively. The refolding of the denatured inclusion body was successful by gradual dilution. Further purification of the refolded protein and soluble crude extract from M9 medium with Ni2+ -NTA argarose resulted a 90% purified PNGase F. The purified protein catalyzed the complete and intact cleavage of N-linked oligosaccharides from various glycoproteins. The efficiency of this cleavage was affected by the substrate status in the reaction system. In summary, we have developed an enzyme production system where PNGase F was over-expressed in recombinant E. coli. This system can provide more than 15 mg/L purified active PNGase F. This purified active PNGase F can be used as tools in analyzing the oligosaccharide structure.
Bacterial Proteins ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Flavobacterium ; enzymology ; genetics ; Glycosylation ; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism

One year survey on the concentrations and monthly or seasonal variations of airborne microbe in Guangzhou city were analysed and studied with JWL-IIB airborne microbial sampler. The results showed that the yearly average airborne microbe content of outdoor was 2, 298 cfu/m3, and that of indoor was 1,792 cfu/m3 in Guangzhou city. The monthly variation range of outdoor airborne microbe was from 1,073 to 4,096 cfu/m3, the highest content was 4,096 cfu/m3 in March, and the lowest content was 1,073 cfu/m3 in October. The outdoor airborne bacteria and fungi counts were the highest in spring, next in summer, lower in winter and the lowest in autumn in the four seasons . The yearly average concentrations of outdoor airborne microbe at the Garbage compression station, the business walk street, the key traffic route, the residential area, the industrial district and the garden were 4, 573, 3, 835, 1, 580, 1,413, 1, 197 and 1, 187 cfu/m3respectively; and Ones of indoor at the key traffic route, the tourist three star-route hotel and the subway station were 2,511, 1,699and 1,167 cfu/m3 respectively . The study on airborne microbe can be used for the research of health prevention and environment control measures in Guangzhou .

OBJECTIVETransfusion transmitted virus (TTV) DNA was detected in serum samples obtained from healthy infants and volunteer blood donors living in Jiujiang city in an attempt to shed light on the prevalence of TTV infection and the transmission route of TTV infection in infants.

METHODSModified untranslated region, polymerase chain reaction (UTR PCR) and N22 PCR were performed to test TTV DNA in serum samples from 86 infants and 58 blood donors.

RESULTSTTV DNA was detected by UTR PCR in 51 (53.5%) infants and 58 (100%) in blood donors, while that tested by N22 PCR was 14 (16.3%) and 22 (37.3%) in infants and blood donors, respectively. Among infants younger than 30 days, 1 - 6 months and 7 - 12 months of age, TTV DNA was detected by UTR PCR and N22 PCR at rates of 0, 33.3%, 95.0% and 0, 7.4%, 30.0%, respectively.

CONCLUSIONThe prevalence rates of TTV DNA detected by UTR PCR were 95% in infants of 7 - 12 months after birth and 100% in healthy blood donors in Jiujiang city. However the results obtained by N22 PCR were much less frequently in the same population. Results showed that significant difference did exist in the prevalence of TTV DNA detected by the two different PCR systems. Age-dependent increase of TTV infection was observed in early childhood, while environmental sources were considered to be the most common route of TTV acquisition as the primary infection in infants. However, the prevalence of TTV in infants of 7 - 12 months was similar to that in healthy adults in the same region.

Base Sequence ; China ; epidemiology ; DNA Virus Infections ; epidemiology ; virology ; DNA, Viral ; chemistry ; genetics ; Humans ; Infant ; Infant, Newborn ; Molecular Sequence Data ; Polymerase Chain Reaction ; Prevalence ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Torque teno virus ; genetics

OBJECTIVE: To study the regulatory mechanism of MS275, a histone deacetylase inhibitor, on the p38 MAPK signaling pathway in rats with convulsion in the developmental stage. METHODS: Thirty-two male rats were randomly divided into four groups: control, pentylenetetrazol (PTZ), PTZ+3 mg/kg MS275, and PTZ+6 mg/kg MS275 (n=8 each). A rat model of convulsion in the developmental stage was prepared by an intraperitoneal injection of PTZ. The rats in the control group were given an injection of normal saline alone. MS275 was given by an intraperitoneal injection at 2 hours before PTZ injection. At 24 hours after successful modeling, 6 rats were taken from each group. Western blot and qRT-PCR were used to measure the protein and mRNA expression of p38, MK2, cAMP response element-binding protein (CREB), and interleukin-6 (IL-6) in the hippocampus. Hematoxylin-eosin (HE) staining was used to observe brain pathological changes. Western blot was used to measure the expression of CD11b as a marker for the activation of microglial cells. RESULTS: Compared with the control group, the PTZ group had significant increases in the mRNA and protein expression of p38, MK2, CREB, and IL-6 (P<0.05). MS275 significantly inhibited the mRNA and protein expression of the above markers in the rats with convulsion in the developmental stage (P<0.05), and 6 mg/kg MS275 had a significantly better inhibitory effect on the mRNA and protein expression of IL-6 and CREB than 3 mg/kg MS275 (P<0.05). HE staining showed that the PTZ group had marked neuron apoptosis, cellular edema, and inflammatory cell infiltration, while MS275 intervention alleviated neuron apoptosis and cellular edema and reduced inflammatory cell infiltration in the rats with convulsion. The PTZ group had a significant increase in the activation of microglial cells, while MS275 significantly inhibited the activation of microglial cells in the rats with convulsion (P<0.05); 6 mg/kg MS275 had a significantly better inhibitory effect than 3 mg/kg MS275 (P<0.05). CONCLUSIONS In rats with convulsion in the developmental stage, the histone deacetylase inhibitor MS275 can inhibit the p38 MAPK signaling pathway, the apoptosis of hippocampal neurons, and the activation of microglial cells and thus reduce inflammatory response and convulsion-induced brain injury in a dose-dependent manner.
Animals ; Male ; Pentylenetetrazole ; Rats ; Rats, Sprague-Dawley ; Seizures ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases