In order to construct a recombinant attenuated Salmonella typhimurium expressing Helicobacter pylori (H. pylori) urease subunit B(UreB), UreB gene fragment amplified by PCR was cloned into the prokaryotic expression vector pTC01 after sequencing, then transformed into attenuated Salmonella typhimurium SL3261 to acquire SL3261/pTC01-UreB. The expression of H. pylori UreB in SL3261 was detected by Western blot. Tewelve weeks after oral immunization of mice with this vaccine, anti-UreB IgA antibodies in mouse intestinal fluid and IgG antibodies in serum were determined by ELISA. IFN-? and IL-10 contents in the supernatant of spleen cell culture were also assessed by ELISA. The results showed that 61kD protein was expressed in SL3261/pTC01-UreB that could be recognized by anti-H. pylori UreB antiserum by Western-blot. The multiple oral immunizations with SL3261/pTC01-UreB could induce significantly H. pylori-specific mucosal IgA response as well as serum IgG responses. Moreover, there was significant increase of IFN-? and IL-10 contents in the supernatant of spleen cell culture in SL3261/pTC01-UreB group. These results suggested that the attenuated Salmonella typhimurium expressing H. pylori UreB may be used as oral vaccine against H. pylori infection. Its effect against H. pylori infection also needs to be further evaluated in animal models.

To investigatethe relationship between p38 MAPK pathway and multidrug resistance of stomach cancer. The activity of p38 MAP kinase in stomach cancer cell SGC7901 and multidrug resistant cell SGC7901/VCR were examined by immunoprecipition after adriamycin treatment SGC7901 for 0, 2, 15, 30 and SGC7901/VCR for 0, 2, 15, 30, 60 minutes,respectively. The results indicated that adriamycin could activate the activity of p38 MAP kinase in gastric cancer cell SGC7901 with time dependency, but could decrease remarkably the activity of p38 MAP kinase in gastric cancer multidrug resistant cell SGC7901/VCR after adriamycin treatment it for 15 minutes. It suggested that adriamycin could affect the activity of p38 MAPK in gastric cancer cell SGC7901 and multidrug resistant cell SGC7901/VCR.

Objective To study the relation between p38 MAPK pathway and multidrug resistance of stomach cancer. Methods The activity of p38 MAPK in gastric cancer cell SGC7901 and multidrug resistant cell SGC7901/VCR was examined by immunoprecipitation after treating with vincristine for 0,2,15,30,60 minutes. Results Vincristine could activate the activity of p38 MAPK in gastric cancer cell SGC7901 and multidrug resistant cell SGC7901/VCR. The difference between them was that in cell SGC7901 vincristine activate the p38 MAPK rapidly whereas in cell SGC7901/VCR slowly. Conclusion Vincristine could affect the activity of p38 MAPK in gastric cancer cell SGC7901 and multidrug resistant cell SGC7901/VCR.

Objective To establish attenuated S. typhimurium expressing Helicobacter pylori (H.pylori) urease subunit B (UreB) and determine whether it could be used as oral vaccine against H.pylori Methods H.pylori (SS1 strain) UreB gene fragment amplified by PCR was cloned into the prokaryotic expression vector PTc 01 after sequencing, then transformed into attenuated S.typhimurium SL3261 to acquire SL3261/PTc 01 UreB. The expression of H.pylori UreB in SL3261 was detected by Western blot. Twelve weeks after oral immunization of mice, antibody responses were evaluated using serum and intestinal fluid by ELISA. In vitro stability of PTc 01 UreB plasmid in S. typhimurium SL3261 was confirmed by growing in Luria Bertani medium to 60 generations. Results The UreB gene fragment amplified by PCR was consistent with the sequence of the H.pylori UreB by sequence analysis. Enzyme digestion revealed that the correct PTc 01 UreB was obtained. Western blot showed that 61kD protein was expressed in SL3261/PTc 01 UreB which could be recognized by anti H.pylori UreB antiserum. Anti UreB IgA antibodies in mouse intestinal fluid and IgG antibodies in serum were determined by ELISA. After 60 generations of continuous culture, the recombinant plasmid PTc 01 UreB was stable in SL3261 and had no obvious toxicity. Conclusion The attenuated Salmonella typhimurium expressing H.pylori UreB may be used as oral vaccine against H.pylori infection.

[Abstract ]Objective:To estimate dental maturation norms of permanent dentition in a group of Nanjing children using Nolla's tech-nique.Methods:549 cases of panoramic radiographs (279 males and 270 females)of Nanjing children at the age range of 3 to 16 years were collected for the study.All permanent mandibular teeth on the left side (except for the third molars)were scored according to Nolla's method,and data were presented in tables and used to plot dental maturation curves for the Nanjing boys and girls.Results:Females were found to be more advanced in the degrees of dental development.Dental maturation is significantly associated with chron-ological age (for males and females:r =0.959,P <0.001 and r =0.953,P <0.001,respectively).The mathematical model of them was established,and then the fitting equation was obtained.Conclusion:The dental maturation curve and chronology of permanent dentition can provide the growth norms of permanent teeth for Nanjing children and facilitate the way for dentists to assess the growing children during diagnosis and treatment planning.

A new group of gastric cancer associated antigens in sera of patients with rectal cancer, benign diseases and normal human were detected by MG series monoclonal antibodies against gastric cancer. The average values of normal persons plus 3 standard deviations was set as the highest limit of normal value. The positive rate were 69.7% (23/33) in sera of patients with rectal cancer, and 4%(1/25) in sera of patients with benign diseases. The values of MG-Ags were decreased to normal level 8-10 days after surgical removal in 5 patients of rectal cancer. The results of test suggest that determination of MG-Ags in serum of patients might be helpful in the diagnosis of rectal carcinoma.

Objective To study the method of gray-scale blood flow imaging and image processing in condition of high frequency ultrasound,and the implementation of the system.Methods On the base of research of scattered signals of red blood cells in high frequency ultrasound,20 MHz ultrasound mechanical and linear scanning probe was used to transmit a number of pulses on a scan line.Pulse-echo subtraction method was used to obtain the blood flow information.At the end,simulated blood vascular was used to conduct flow imaging,and the obtained images were analyzed.Results Experiment results showed that clear blood flow images were obtained using this system.The noise from perivascular tissue could be filtered and the signals from blood flow could be enhanced after image processing.Conclusions In the detection of superficial blood vessel,blood flow signals can be obtained even using single pulse emitting via high frequency ultrasound.The blood flow imaging system can be implemented after image processing.

OBJECTIVETo establish attenuated Salmonella typhimurium producing Helicobacter pylori (H. pylori) urease subunit B (UreB) and determine whether it could be used as an oral vaccine against H. pylori.

METHODSH. pylori (SS1 strain) UreB gene fragment amplified by PCR was cloned into the prokaryotic expression vector pTC01 after sequencing, and then transformed into attenuated Salmonella typhimurium SL3261 to acquire SL3261/pTC01-UreB. The expression of H. pylori UreB in SL3261 was detected by Western blot. Twelve weeks after oral immunization of mice, antibody responses were evaluated using serum and intestinal fluid by ELISA assay. Interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) in the supernatant of spleen cells culture were also assessed by ELISA. In vitro stability of pTC01-UreB plasmid in SL3261 was confirmed by growing in Luria Broth (LB) medium to 80 generations.

RESULTSThe UreB gene fragment amplified by PCR was consistent with the sequence of the H. pylori UreB as evidenced by sequence analysis. Enzyme digestion revealed that the correct pTC01-UreB was obtained. Western blot showed that a 61kDa protein was expressed in SL3261/pTC01-UreB, which could be recognized by anti-H. pylori UreB antiserum. After 80 generations of continuous culture, the recombinant plasmid pTC01-UreB was stable in SL3261 and had no obvious toxicity. Multiple oral immunizations with SL3261/pTC01-UreB could significantly induce H. pylori-specific mucosal IgA response as well as serum IgG response. Moreover, there were significant increases of IFN-gamma and IL-10 in the SL3261/pTC01-UreB group. Finally, no obvious side effects for mice and no change in gastric inflammation were observed.

CONCLUSIONAttenuated Salmonella typhimurium expressing H. pylori UreB may be used as oral vaccine against H. pylori infection.

Administration, Oral ; Animals ; Antibodies, Bacterial ; blood ; Bacterial Vaccines ; blood ; immunology ; Female ; Helicobacter Infections ; prevention & control ; Helicobacter pylori ; immunology ; Immunization ; Interferon-gamma ; biosynthesis ; Interleukin-10 ; biosynthesis ; Mice ; Mice, Inbred BALB C ; Plasmids ; Protein Subunits ; Salmonella typhimurium ; genetics ; Urease ; immunology ; Vaccines, Attenuated ; immunology ; Vaccines, Synthetic ; immunology

Colorectal cancer (CRC) belongs to multiple diseases of traditional Chinese medicine (TCM).CRC can exhibit sophisticated clinical manifestations and various TCM syndromes (Zheng).Modem studies have revealed that syndromes (Zheng) of CRC are related to pathological characteristics,cell types,staging and prognosis.Surgery,chemotherapy and other modem treatments can influence the manifestations of syndromes (Zheng) of CRC.Modern TCM doctors have extensively studied the biomedical basis of syndromes (Zheng) of CRC,including accumulation of damp-heat (Shi-Re-Yun-Jie),blood stasis (Yu-Xue-Nei-Zu),spleen (Pi) deficiency (Pi-Qi-Kui-Xu),deficiency of qi and blood (Qi-Xue-Liang-Xu),deficiency of yin of liver (Gan) and kidney (Shen) (Gan-Shen-Yin-Xu) and other syndromes (Zheng).These investigations may contribute to the objective and standardization study of syndromes (Zheng) in CRC.