Objective To investigate the change of immunologic function of red blood cell (RBC) in patients with chronic gastritis. Methods The immunologic function of RBC in 88 cases of chronic gastritis was dynamically detected with yeast rosette method. Results The results showed that RBC-C3b receptor rosette (RBC-C3b-RR) rate and RBC immune adherence enhance factor (RFER) rate were significantly lower (P＜0.05) and RBC immune complex rosette (RBC-ICR) rate and RBC immune adherence inhibitor factor (RFIR) were significantly higher (P＜0.05) in the patients with atrophic gastritis than those in control group. In the patients with superficial gastritis, the difference had no distinct significance (P＞0.05). Conclusion The immunologic function of red blood cell (RBC)measurement plays an important role in differential diagnosis and prognosis evaluation of chronic gastritis.

[Objective]To observe the growth pattern of oligodendrocyte precursor cells(OPCs) in the primary culture from neonatal rat cerebral cortices and study the methods of cell culture and isolation in order to obtain purified OPCs for the experiments of cell transplantation.[Method]The mixed glial cells from the cerebral cortices of 48-hour-old Sprague-Dawley(SD) rats were cultured in vitro.Until 9-10 days in vitro OPCs were isolated and purified by the shaking process and differential adhesion,and then continued OPCs culture in the defined medium.The growth pattern of OPCs in vitro was investigated by contrast phase microscopy and OPC s were further identified with the immunocytochemical techniques.[Result]The distinct stratification of OPCs and astrocytes developed around 9-10 days in primary culture.At this point,the OPCs scattered on the top of the monolayer astroctyes and the soma of most OPCs typically appeared oval or round with two or three processes.The purity of the isolated OPCs reached 95% and these OPCs further developed into the mature oligdendrocytes which were immunoreactive to the specific antigen of oligodendrocyte lineage cells,Oligo2.[Conclusion]OPCs separated from the cerebral cortices of neonatal SD rats can be maintained as immature precursor cells in culture,and OPCs are able to be purified by shaking and differential adhesion under the condition of appropriate cell stratification.

Objective To investigate the effects of transplantation of oligodendrocyte precursor cells (OPCs) on axonal myelination after spinal cord injury in a rat model. Methods A rat model of spinal cord injury at the tenth thoracic vertebral level (T10) was produced by Allen weight-drop impact method. OPCs implantation was performed at the subacute stage of spinal cord injury. Effects of OPCs transplantation on axonal myelination after spinal cord injury were evaluated by HE staining, immunohistochemistry, myelin staining and transmission electron microscopy. Results The implanted cells were still observed in lesioned segments of spinal cord eight weeks after transplantation. The results of HE staining clearly showed better structure of spinal cord in OPCs-transplanted group than that of control group.Myelin staining also demonstrated that the amount of myelin in white matter of lesioned cord in the OPCs-transplanted group (7 802.42 ± 1085.58) was higher than that of the control group (5 055.98 ± 916.74)(P ＜0.01 ). Expression of myelin basic protein (MBP) was significantly increased in the OPCs-trans-planted group (8 544.44 ±812.78) as compared with that of the control group (5 243.83 ±808.27)(P＜0.01). Moreover, transmission electron microscopy further confirmed the improvement of micro-structure of myelination in OPCs-treated rats. Conclusion OPCs transplantation can improve axonal myelination in rat with spinal cord injury.

Objective To clone the cheA and cheY genes of Helicobacter pylori for construction of their prokaryotic expression systems, and to establish chemotactic model in vitro of H. pylori for determing chemotaxis-inducing substances and to understand the effects of specific antibody and closantel on inhibiting chemotactic behavior of the microbe. Methods The segments of entire cheA and cheY genes were amplified by PGR and then sequenced after T-A cloning. Prokaryotic expression systems of the genes were subsequent-ly constructed. SDS-PAGE plus Bio-Rad Gel Image Analyzer were used to examine the expression of target recombinant proteins rCheA and rCheY, and Ni-NTA affinity chromatography was performed to extract rCheA and rCheY. Rabbits were immunized with rCheA and rCheY to obtain antisera and IgG in each of the anti-sera was extracted by saturated ammonium sulfate precipitation and DEAE-32 ion exchange chromatography. Immunodiffusion assay was performed to measure the titers of antisera and their IgGs. Chemotactic model in vitro of H. pylori based on hard-agar plus method was established to determine the chemotaxis-inducing effects of eleven candidate substances. Simultaneously, the effects of rCheA-lgG and closantel sodium on blocking the bacterial chemotactic behavior were also observed. Results The segments with expected sizes of cheA and cheY genes were obtained by PCR, and their nucleotide and putative amino acid sequences were 100% idenities to the reports. The constructed prokaryotic systems could efficiently express rCheA and rCheY. The two rabbit antisera and IgG aginst rCheA and rCheY had 1 : 4 and 1 : 2 immunodiffusion titers, respectively. Hydrochloric acid, sulfuric acid and acetic acid were able to induce chemotactic movement of H. pylori. Both rCheA-IgG and closantel sodium with certain concentrations could weaken the chemotactic ability of H. pylori(P<0.05). Conclusion The prokaryotic expression systems of H. pylori cheA and cheY genes were successfully generated in this study. Hydrogen ion (H~+) is the inducer for chemotaxis of H. py-lori. rCheA-IgG, as well as closantel sodium can inhibit H~+-induced chemotaxis of H. pylori.

Objective To determine the effect of cheA gene of Helicobacter pylori in the bacterial chemotaxis in vitro and colonization in vivo. Methods The entire cheA and cheY genes were amplified and cloned from genomic DNA of H. pylori NCTC11637 strain. Subsequently, the prokaryotic expression systems of cheA and cheY genes were generated and the target recombinant proteins rCheA and rCheY were extracted by Ni-NTA affinity chromatography. Rabbits were immunized with either rCheA or rCheY for obtaining antisera, and rCheA-IgG and rCheY-IgG in the antisera were prepared using ammonium sulfate precipitation plus DEAE-52 column chromatography. A suicide plasmid of cheA gene was constructed and then a cheA gene knock-out mutant ( cheA - ) was generated based on homologous recombinant exchange using the suicide plasmid. The cheA- mutant was identified using PCR and sequencing. The phosphorylation levels of CheA and CheY molecules of cheA - and wild-type strain were determined by using rCheA-IgG and rCheY-IgG anchoring the target proteins and protein phosphorylation detection kit. The differences of chemotaxis in vitro and colonization in vivo between cheA- mutant and wild-type strain were compared using chemotactic model and BALB/c infection model of H. pylori. Results The cheA gene knock-out in genome of cheA- mutant was confirmed by the results of PCR and sequencing. After treated with 0. 001-0. 1 mol/L HCI for 10 min, the phosphorylation levels of CheA and CheY molecules of wild-type strain were rapidly descended from ( 59.6 ±11.5) μmol and (55.5 ± 10.2) μmol to ( 10.8 ± 2.6) and (5. 5 ± 1.2) μmol (P ＜ 0.05 ), while the phosphorylation of CheY molecule of cheA - mutant was no markedly changed with a persistent lower level ( P ＞0.05). The diameters [(10-20) ± (2-3) mm] of chemotactic aggregative rings of cheA- mutant were significantly less than those [(16-24) ± (2-3)mm] of wild-type strain (P ＜0.05). The positive isolation rate (90%) of H. pylori in gastric biopsy specimens of mice that infected with wild-type strain was remarkably higher than that (40%) of mice that infected with cheA- mutant (P ＜0.05). The result of fluorescence quantitative was also showed that the numbers (6.3 × 103 ±2.1 × 103 copies/mg) of H. pylori in gastric biopsy specimens of wild-type strain infected mice were significantly larger than those (8.3 × 101 ±3. 1 × 101 copies/mg) in gastric biopsy specimens ofcheA- mutant infected mice (P＜0.05). Conclusion The cheA gene of H. pylori has an important role in the bacterial chemotaxis in vitro and colonization in vivo.

ObjectiveTo optimize the condition of multiple fluorescence quantitative PCR and establish a new assay of four human herpes virus (HHV) detected by AllGlo quadruple fluorescence quantitative PCR.MethodsFour HHV including HSV-1,HSV-2,EBV and CMV were identified by sequence analysing the qualitative PCR production.Furthermore,they were quantitatively detected by AllGlo and TaqMan multiple fluorescence quantitative PCR respectively.ResultsBoth the positive rate and specificity of AllGlo and TaqMan in detecting single HHV achieved 100％.And AllGlo single fluorescence quantitative PCR prevailed over TaqMan's by Ct of 1-3.Four HHV can be simultaneously detected by AllGlo quadruple fluorescence quantitative PCR,comparing to the only two by TaqMan.ConclusionAllGlo fluorescence quantitative PCR assay allows a higher throughput,sensitivity and specificity than TaqMan in detection and thus provides a board prospect.

Objective To evaluate the application of fluorescence PCR in detecting ureA gene of Helicobacter pylori(HP)in feces.Methods Fluorescence PCR was used to detect ureA gene of HP in feces from 50 patients,including 23 confirmed by gastric biopsy urease test and histological staining.Bacterium culture and serum antibody detection were also performed, and chi-square test was used to compare the sensitivity,specificity,positive predictive value and negative predictive value among three methods.Results The sensitivity,specificity,positive predictive value and negative predictive value of fluorescence PCR were 1.00,0.96,96.O%and 100.0%,while those for HP culture were 0.78,1.00,100.0%and 84.0%,and thee for serum antibody detection Was 0.96,0.74,76.O%and 95.0%.There were significant differences in sensitivity and negative predictive value between PCR and bacterium culture (X2=5.60 and 4.44,P<0.05),and significant differences in specificity and positive predictive value between PCR and serum antibody detection(X2=5.28 and 4.08,P<0.05).Conclusion ureA gene detection in feces by fluorescence PCR is of value for the diagnosis of HP infection.

Marine traditional Chinese medicine (MTCM) is an important part of Chinese medicine (CM),there are some differences in understanding of the current literature and the extension of the connotation of marine medicine,which leads to the definition dispute of MTCM,hindering clinical application and further development of MTCM.In this study,we explored the concept of MTCM in literature,discussed the attributes of ocean marine CM,summed up the differences between the land CM and MTCM over variety characteristics,effect of drug composition characteristics and biological activity characteristics,and discussed the connotation and extension of MTCM from three aspects of theoretical basis and the effect and source of the drug,leading to the formation of the narrow and broad concept of MTCM.The five kinds of disputes in the definition of MTCM were discriminated according to the concept and connotation,which provided a theoretical basis for the definition and the research of MTCM.Moreover,we also defined the English translation and its abbreviation as Marine Traditional Chinese Medicine (MTCM).

Objective To investigate the role of cytokines genes expression of liver sinusoidal endothelial cells in liver damage by sepsis.Methods Septic mice models were established with cecal ligation and perforation (CLP), while sham operation group received the same procedure exclusive CLP. The genes expression of TNF?, IL 1? and IL 6 in liver sinusoidal endothelial cells were assessed by RT PCR. Results A significant increase of TNF?, IL 1? genes expression was observed at 3h, and a slight decline at 12h after operation, but still significantly higher than that in the sham group; while IL 6 gene expression showed signficantly higher at 3h and remained at the high level at 12h. Conclusions Liver sinusoidal endothelial cell is an important source of cytokine production in mice with sepsis.

Objective To investigate the effects of inhibiting ubiquitin proteasome pathway(UPP) on proliferation of gastric carcinoma cells and the possible mechanism was discussed. Methods The gastric carcinoma cell strain SGC 7901 was treated with MG 132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay. Cell cycle and apoptosis were detected by flow cytometry(FCM). DNA fragment analysis was used for confirming the presence of apoptosis. The activity of telomerase was examined by TRAP PCR ELISA. Expression of p27kip1 was detected by immunocytochemical technique. Results MG 132 had great inhibitory effect on the growth of SGC 7901 cells. The FCM analysis showed that the ratio of G0/G1 phase of control group was (46.3?4.1)%, the ratio of G0/G1 phase of SGC 7901 cells treated with MG 132 increased to (72.1?5.0)% ( P