Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 24
Filter
1.
Chinese Journal of Orthopaedic Trauma ; (12): 70-76, 2023.
Article in Chinese | WPRIM | ID: wpr-992683

ABSTRACT

Objective:To establish a rat model of transverse tibial bone transfer on the diabetic foot.Methods:A diabetic model was created by intraperitoneal injection of streptozotocin into 40 SD rats after 5 weeks of high-fat feeding, taking random blood glucose ≥16.7 mmol/L as the criterion for successful modeling. Changes in body weight, food intake, water intake, faecal output and blood glucose were monitored every week after the acclimatization period until random blood glucose ≥16.7 mmol/L was observed for 3 running weeks. After the blood glucose was stabilized, 34 surviving diabetic rats were divided into 2 groups using a random number table. In the experimental group, a transverse transfer outer frame was installed and transverse tibial bone transfer performed after removal of the skin over the dorsal foot; in the control group, a transverse transfer outer frame was installed and the skin over the dorsal foot removed but no transverse tibial bone transfer performed. The wound changes were recorded on the 1, 5, 10, 15 and 20 days after installation of the transverse transfer outer frame. After completion of transverse tibial bone transfer (24 days after modeling of transverse tibial bone transfer), one rat was randomly chosen from each of the 2 groups for angiography while the rest rats were sacrificed for observation of the skin changes on the lower limbs. The densitometric values of CD31 immunohistochemical staining were compared between the 2 groups.Results:The random blood glucose increased from (6.89±1.03) mmol/L before modeling to (25.91±6.42) mmol/L at the last test, keeping at ≥16.7 mmol/L for 3 running weeks. The percentages of ulcer healing of the foot dorsal wound in the experimental group on days 5, 10, and 20 were significantly higher than those in the control group ( P<0.05). Gross observation and angiography found more abundant vessels in the lower extremity on the side of transverse transfer in the experimental group. The immunohistochemical staining of the lower limb skin tissue for CD31 found an AOD value of 0.60±0.23 in the experimental group, significantly higher than that in the control group (0.37±0.13) ( t=3.722, P=0.001). Conclusion:A rat model of transverse tibial bone transfer has been successfully established through installation of a self-designed transverse transfer outer frame on the basis of a successful rat model of diabetic foot, characterized by improved microcirculation in the lower limbs.

2.
Chinese Journal of Orthopaedic Trauma ; (12): 339-344, 2022.
Article in Chinese | WPRIM | ID: wpr-932335

ABSTRACT

Objective:To explore the effects of 3 osteotomy methods on the bone healing in Ilizarov tibial bone transport.Methods:The data of 93 patients were retrospectively reviewed who had been treated by Ilizarov single-segment tibial bone transport at Department of Hand Surgery, The Second Hospital of Tangshan from December 2003 to April 2019. Minimally invasive osteotomy was performed in 16 patients [group A: 16 males with an age of (37.1±8.3) years; 5 cases of type Ⅱ and 11 ones of type Ⅲ by Gustilo classification], subperiosteal saw osteotomy in 57 patients [group B: 47 males and 10 females with an age of (39.1±11.8) years; 17 cases of type Ⅱ and 40 ones of type Ⅲ by Gustilo classification] and extraperiosteal wire saw osteotomy in 20 patients [group C: 19 males and one female with an age of (37.7±11.2) years; 18 cases of type Ⅱ and 2 cases of type Ⅲ by Gustilo classification]. The 3 groups were compared in terms of the bone healing index and the Association for the Study and Application of the Method of Ilizarov (ASAMI) functional scores.Results:The 3 groups were comparable because there was no significant difference in the preoperative general data between them ( P>0.05). All the patients were followed up for 19 to 50 months (average, 27.4 months). All patients achieved bony healing, and their associated complications were cured after corresponding treatments. There were no significant differences in the bone healing index between the 3 groups [(53.09±21.88) d/cm for group A, (59.97±33.29) d/cm for group B and (46.20±14.11) d/cm for group C] ( P>0.05). There were no significant differences either in the good to excellent rate by the ASAMI functional scores between the 3 groups (87.5% for group A, 89.5% for group B and 90.0% for group C) ( P> 0.05). Conclusion:All the 3 osteotomy methods may achieve good bony union, leading to similar bone healing indexes and postoperative functional scores.

3.
Chinese Journal of Medical Aesthetics and Cosmetology ; (6): 413-417, 2022.
Article in Chinese | WPRIM | ID: wpr-958748

ABSTRACT

Objective:To explore the risk factors of skin flap pigmentation after repairing skin and soft tissue defects of extremities using cutaneous neurovascular flap and to give some suggestions for prevention and treatment.Methods:From January 2013 to March 2020, 160 cases of extremities cutaneous nerve vascular flap with survival in Tangshan Second Hospital were retrospectively studied. According to the occurrence of pigmentation, they were divided into two groups: Group A (pigmentation group) and Group B (non-pigmentation group). The observation indexes included sex, age, injury cause, defect size, complete debridement, anastomosis of skin flap, sunscreen measures and postoperative infection. First, univariate analysis was carried out to screen the influencing factors of pigmentation in the cutaneous neurovascular flap, and then Logistic regression was used for multivariate analysis to screen the risk factors.Results:The postoperative follow-up time was 12 to 24 months, with an average of 17.9 months. A total of 29 patients (18.1%) had skin flap pigmentation. Univariate analysis showed that there was no significant difference in sex, age, cause of injury and defect area between the two groups ( P>0.05). Further multivariate analysis showed that incomplete debridement, lack of venous anastomosis, failure to take sunscreen measures and postoperative infection were the risk factors of pigmentation of cutaneous neurovascular flap ( OR=0.310, 0.335, 0.355、5.878, 95% CI=0.112-0.863, 0.115-0.975, 0.133-0.949, 2.069-16.697, P<0.05). Conclusions:Incomplete debridement, lack of venous anastomosis, failure to take sunscreen measures and postoperative infection are the risk factors resulting in pigmentation of neurovascular flap. It is important to perform prevention to reduce the incidence of pigmentation.

4.
Chinese Journal of Pancreatology ; (6): 116-119, 2010.
Article in Chinese | WPRIM | ID: wpr-390048

ABSTRACT

Objective To investigate the changes of extraceUular regulated kinase 1/2 (ERK1/2) phosphorylation and assess the effects of blocking the ERK1/2 phosphorylation on rats with acute necrotizing pancreatitis (ANP). Methods The ANP model was induced by retrograde injection of 5% sodium tanrocholate into the biliary and pancreatic duct. 5 rats were treated as normal control. Other 75 Sprague-Dawley (SD) rots were randomly divided into sham operations(SO) group (n =25), ANP group (n =25) and PD98059 group (n =25). The rats were sacrificed at 15 min, 30 min, 1 h, 3 h and 6 h after ANP induction, the blood and pancreatic sample were taken. Pathological changes of pancreas were observed with light microscope and scored. The serum level of TNF-α and IL-1β was determined by ELISA. MPO activities in pancreas were measured by enzyme chemistry assay. Western blotting was performed to determine the phosphorylations of ERK1/2 in the pancreas homogenates. Results There was no significant pathologic changes in rats of SO group;but significant injuries occurred in ANP group, the pathologic score at 3 h was 9.9 ± 0.4;the extent of injuries attenuated in PD98059 group, the pathologic score at 3 h was 4.0 ± 0.4 (P < 0.05). The serum levels of TNF-α at 3 h in SO, ANP and PD98059 groups were (65.8 ± 20.5) pg/ml, (286.5 ± 50.3) pg/ml, (180.4±32.9)pg/ml, respectively;the serum levels of IL-1β at 3 h in SO, ANP and PD98059groups were (85.8 ± 25.5) pg/ml, (293.8 ± 46.3) pg/ml, (200. 5 ± 33.6) pg/ml, respectively;MPOactivities in pancreas were (0. 19 ± 0.02)U/g, (0.61±0.05)U/g, (0.52±0.03) U/g, and the values in ANP and PD98059 groups were significantly higher than those in SO group, while the values in PD98059 group were significantly lower than those in ANP group (P < 0.01). The expression of ERK1/2 phosphorylation in normal pancreas was 1100 ± 141, the expressions of ERK1/2 phosphorylation in ANP group at 15 min, 30 min were 5300 ± 486, 5621 ± 384, respectively;the expressions began to decrease 1 h later and returned the similar level as SO group at 6 h;the expressions of ERK1/2 phosphorylation in PD98059 group at 15 min, 30 rain were 4200 ± 370, 3600 ± 290, respectively;which were signifieanfly lower than those in ANP group (all P value < 0. 01). Conclusions The ERK1/2 signal transduetion pathway plays an important role in the pathogenesis of ANP. Inhibition of ERK1/2 phosphorylation by PD98059 may decrease the production of IL-1β, TNF-α and pancreatic MPO, attenuate the extent of pancreatic pathologic injuries.

5.
Chinese Journal of Pancreatology ; (6): 305-307, 2008.
Article in Chinese | WPRIM | ID: wpr-397717

ABSTRACT

Objective To investigate the expression of Fascin mRNA and its protein in human pancreatic carcinoma cell line and pancreatic cancer tissues and to explore the relationship between the expression of Fascin protein and the clinicopathologic parameters. Methods The expression of Fascin mRNA in pancreatic cancer cell lines (SW1990, Patu8988, BxPC3, cfPAC1) were measured by RT-PCR and the expression of Fascin protein in 54 samples of pancreatic career tissues and 42 adjacent normal pancreatic tissues was detected by immunohistochemical method. Results The expression of Fascin mRNA was confirmed in 3 of 4 pancreatic cancer cell lines (SW1990, Patu8988, cfPAC1), but not in the cell line of BxPC3; the rate of positive expression of Fascin protein in pancreatic cancer tissues was 64.81% (35/54) and there was no positive expression in adjacent normal pancreas tissues; the expression of Fascin protein correlated with the differentiation degree (P < 0.01) and with the lymphatic metastasis of the carcinoma (P <0.05), but not with the size and distant metastasis of the carcinoma (P > 0. 05). Conclusions Fascin protein was highly positively expressed in pancreatic cancer tissues, and the expression of Fascin protein may help diagnose pancreatic, carcinoma and predict the malignant degree.

6.
Journal of Third Military Medical University ; (24)2003.
Article in Chinese | WPRIM | ID: wpr-558614

ABSTRACT

Objective To investigate the effect of esophageal mucosal acid exposure on visceral sensation of patients with non-erosive gastroesophageal reflux disease (NERD) and to evaluate the role of visceral hypersensitivity in NERD pathogenesis. Methods We recruited 21 NERD patients and 10 normal healthy volunteers. Mechanical distentions stimulation and acid perfusion through esophagus were performed using the balloon-affixed and polyvinyl multilumen catheter. Esophageal visceral perception thresholds were examined before and after acid perfusion with esophageal balloon distention by means of a computer-controlled barostat. Results As compared with healthy subjects, NERD patients demonstrated significantly lower initial perception threshold and maximally tolerated pain threshold (P

7.
Chinese Medical Journal ; (24): 1205-1208, 2002.
Article in English | WPRIM | ID: wpr-340356

ABSTRACT

<p><b>OBJECTIVE</b>To determine the efficacy of adenovirus mediated suicide gene transduction combined with prodrug 5-fluorocytosine (5FC) as a therapeutic protocol for pancreatic cancer.</p><p><b>METHODS</b>Cytosine Deaminase(CD) gene was cloned into pAdTrack-CMV-CD, pAdTrack-CMV-CD and pAdEasy-1 were recombined in bacteria. The newly recombined adenovirus (Ad)-CD containing green fluorescent protein (GFP) were packaged and propagated in 293 cells and purified by cesium chloride gradient centrifugation. Human pancreatic carcinoma cell line-Patu8988 was infected with this virus, then 5FC was added. XTT assay was used to estimate relative numbers of viable cells. In vivo model of pancreatic cancer was established by injecting 1.0 x 10(7) Patu8988 cells subcutaneously in Balb/c nude mice. When tumors were palpable, Ad-CD was injected into each tumor and 5FC was administered.</p><p><b>RESULTS</b>Positive clones were selected using endonuclease to digest the recombinants and the concentration of viral liquids containing the CD gene was 2 x 10(11) pfu /ml. Significant cytotoxic activity as shown for 5FC in the CD gene transduced 8988 cell line, while little effect was found in the nontransduced pancreatic carcinoma cells. Antitumor effect was observed in Patu8988 xenograft nude mice with in situ CD gene transduction.</p><p><b>CONCLUSIONS</b>CD gene mediated by adenovirus has high infectivity and may be useful for gene therapy in pancreatic carcinoma. These data demonstrate the use of an enzyme prodrug strategy in experimental pancreatic cancer.</p>


Subject(s)
Animals , Humans , Mice , Adenoviridae , Genetics , Cytosine Deaminase , Gene Transfer Techniques , Genetic Therapy , Genetic Vectors , Mice, Inbred BALB C , Nucleoside Deaminases , Genetics , Pancreatic Neoplasms , Therapeutics
8.
Chinese Journal of Digestion ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-682631

ABSTRACT

Objective To investigate characteristics and alternation of cerebral evoked potentials (CEP) response to esophageal mucosal acid exposure and distention in patients with non-erosive gastro-oesoph- ageal reflux disease (NERD) and in healthy subjects,and to study the mechanism of visceral hypersensitivity in NERD.Methods Twenty-one NERD patients and 10 volunteers were recruited.Mechanical distention stimulation and acid perfusion of the esophagus were performed using the balloon-affixed and polyvinyl multi- lumen catheter.First,maximally tolerated pain thresholds of all subjects were recorded,then esophageal mechanical stimulation with a 75% of maximal tolerated intensity and a frequency of 0.2 Hz was performed altogether 64 times by means of a computer-controlled barostat.The alternation of esophageal CEP was recorded before and after acid perfusion with a multichannel international 10-20 system of electroencephalography. Experimental data was analyzed by student's t-test and one way analysis of variance.Results Esophageal mu- cosal distention may evoke recognizable and reproducible and multi-peak CEP.The latencies for N1,P1 and N2 in volunteers were (246?77),(388?84)and (502?78) ms,CEP morphology of NERD patients was charac- terized by randomly distributed patterns,and the latencies for N1 ,P1 and N2 were (192?46),(293?76) and (440?79)ms,significantly shorter for mechanical stimulation compared with those of control group respectively (all P value

9.
Chinese Journal of Digestion ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-682513

ABSTRACT

Objective To construct nucleic acid vaccine encoding Helicobacter pylori (H. pylori) hpaA gene. Methods The genomic DNA of the standard H. pylori strain 17874 was isolated. hpaA gene fragment was amplified by polymerase chain reaction (PCR) and cloned into pUCmT vector. The sequence of the amplified hpaA gene was assayed, and then cloned into the eukaryotic expression vector pIRES through a series of enzyme digestion and ligation reactions. The recombinant plasmid was transformed into competent Escherichia coli cells DH5?, and the positive clones were screened by PCR reaction and restriction enzyme digestion . Recombinant pIRES hpaA was transfected into COS 7 cells using Lipofectamine TM 2000, and the immunogenicity of expressed HpaA protein was detected with SDS PAGE and Western blot. Results The 750 base pair hpaA gene fragment was amplified from the genomic DNA and was consistant with the sequence of the H. pylori hpaA by sequence analysis. PCR and restriction enzyme digestion results revealed that the H. pylori hpaA gene was inserted into the eukaryotic expression vector pIRES, suggesting the nucleic acid vaccine pIRES hpaA was successfully constructed. The specific protein strip of HpaA expressed by pIRES hpaA was detected with Western blot. Conclusion The nucleic acid vaccine pIRES hpaA was successfully constructed which may help the further investigation towards the immune action of the nucleic acid vaccine.

10.
Chinese Journal of Digestion ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-574753

ABSTRACT

Objective To express the recombinant human canstatin protein, and to examine its biological activity. Methods Canstatin cDNA was cut off from the plasmid pUCm-T/canstatin with restriction enzymes BamHⅠ and Hind Ⅲ. The cDNA fragment was then ligated into the correspondence sites of plasmid pET-22b(+) by T4 DNA ligation enzyme and transformed into E.coli BL21 which was induced to express proteins with isopropyl-1-thio-b-dgalactopyranoside (IPTG). The expressed proteins were analyzed by SDS-PAGE and purified through Ni-NTA column affinity chromatography. Chick chorioallantoic membrane (CAM) assay was performed to determine the activity of the recombinant protein. Results Canstatin cDNA from pUCm-T showed one clear objective DNA band with electrophoresis. Seven of positive colonies were selected and identified by restriction enzyme analysis with BamH Ⅰ and Hind Ⅲ. Electrophoresis revealed that all selected colonies had two specific bands,one near the location of primary plamid,the other near that of objective gene fragment. After IPTG induction, there was a new protein band about 24 000 on SDS-PAGE.The induced product over total bacterial proteins in 1,2, 3. and 4 hours after induction was 18. 2%, 18. 8%, 23.0% and 23.4%, respectively, by densitometry examination. CAM assay demonstrated that the recombinant canstatin protein significantly inhibited the embryonic neovascularization in a dose-dependent manner. Conclusion The prokaryotic expression vector of human canstatin gene has been successfully constructed, laying the foundation for further clinical study.

11.
Chinese Journal of Digestion ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-573232

ABSTRACT

Objective To explore the potential role of cytokine-induced neutrophil chemoattractant (CINC) and monocyte chemoattractant protein-1/JE (MCP-1/JE) in the pathogenesis of early acute pancreatitis(AP). Methods Acute edematous pancreatitis(AEP) and acute necrotizing pancreatitis(ANP) were induced by retrograde infusion of 0.5% or 5% sodium taurocholate into the biliary pancreatic duct. The activity of serum amylase and the level of serum calcium were studied, wet to weight ratio,the activity of myeloperoxidase (MPO) and pathological changes of pancreas were observed. The expression of gene and protein of CINC and MCP-1/JE in pancreatic constitution were detected. Results Compared with that in controls,pancreatic acinar cells were found to express CINC and MCP-1/JE protein in certain intensity. With the induction and the progress of AP,the expression of the activity of MPO,CINC and MCP-1/JE gene in pancreatic parenchyma were all significantly increased ( P

12.
Chinese Journal of Practical Internal Medicine ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-557176

ABSTRACT

Objective To study clinical characteristics and prognosis of patients with liver injury of acute pancreatitis.Methods 290 cases of acute pancreatitis admitted between January 2001 to October 2003 were reviewed and analyzed retrospectively.Results Through comparing different types of AP with liver injury and C-reactive protein (CRP) changes,it was found that the more severe AP was,the more significant liver injury was;and liver injury of severe AP had some connection with CRP(.P.

13.
Medical Journal of Chinese People's Liberation Army ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-556846

ABSTRACT

Objective To clone human canstatin gene, construct its prokaryotic expression vector, express and purify its recombinant protein. Methods The total RNA was extracted from human placenta tissues. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and sequenced. Then the confirmed canstatin cDNA was cloned into plasmid pET-22b(+) and then transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b-Dgalactopyranoside (IPTG). The expression of protein was analyzed through SDS-PAGE. Cells after induced 3 hours by IPTG were harvested, sonicated briefly and the proteins were purified through affinity chromatography. Results (1)The extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis and the concentration was 1.8 g/L. (2)The target sequences were specifically amplified through RT-PCR. (3)The purified RT-PCR product was cloned into pUCm-T vector and then sequenced, demonstrating to be the same as that of canstatin gene in GenBank. (4) the expression vector pET-22b(+) was constructed and verified by the method of BamH Ⅰ and Hind Ⅲ digestion. (5) After IPTG induction, there was a new protein band about Mr 24kD on SDS-PAGE. The percent expressed product over total bacterial proteins after 1, 2, 3, and 4 hours of induction was 18.2%, 18.8%, 23.0% and 23.4%, respectively, estimated by densitometry. (6)After affinity chromatography, SDS-PAGE showed only one clear band existed in 125 mmol/L or 250 mmol/L imidizone elution. Conclusion Human canstatin gene has been successfully cloned and its prokaryotic expression vector has also been successfully constructed. Further more, purified recombinant proteins are obtained through affinity chromatography, laying foundation for further study of its clinical application.

14.
Medical Journal of Chinese People's Liberation Army ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-555305

ABSTRACT

Objective To construct a recombinant live attenuated Salmonella typhimurium nucleic acid vaccine carrying Helicobacter pylori ( H. pylori ) hpaA gene and to detect its immunogenicity. Methods hpaA gene fragment was amplified by polymerase chain reaction (PCR) from the genomic DNA of the standard H. pylori strain and cloned into pUCmT vector. DNA sequence of the amplified hpaA gene was assayed,then cloned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions. The recombinant plasmid was used to transform Escherichia coli DH5?,and the positive clones were screened with PCR reaction and restriction enzyme digestion. Then,the recombinant pIRES-hpaA was transformed to LB5000,then the recombinant plasmid was transformed to SL7207,and the recombinant strain was passaged repeatedly. Finally,recombinant pIRES-hpaA was transfected to COS-7 cells with Lipofectamine TM 2000. The immunogenicity of expressed hpaA protein was determined with SDS-PAGE Western blot. Results The hpaA gene fragment of the 750 base pair was amplified from the genomic DNA,and it was consistent with the sequence of the H. pylori hpaA by sequence analysis. It was confirmed by PCR and restriction enzyme digestion that Helicobacter pylori hpaA gene was inserted into the eukaryotic expression vector pIRES and a stable recombinant live attenuated Salmonella typhimurium nucleic acid vaccine carrying Helicobacter pylori hpaA gene was successfully constructed,and specific strip of hpaA expressed by pIRES-hpaA could be detected with Western blotting. Conclusion The recombinant attenuated Salmonella typhimurium nucleic acid vaccine strain expressing HpaA protein with immunogenicity was constructed,it might be helpful for further investigation concerning the immune action of the nucleic acid vaccine in vivo .

15.
Medical Journal of Chinese People's Liberation Army ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-555300

ABSTRACT

Objective To explore the potential role of the expression of cytokine-induced neutrophil chemoattractant (CINC)gene in the pathogenesis of acute lung injury(ALI)in early severe acute pancreatitis(SAP). Methods SAP was induced by retrograde injection of 5% sodium taurocholate into the bili-pancreatic duct. Wet to weight ratio and myeloperoxidase(MPO) content of lungs were measured. Pathological changes in the pancreas and lungs were observed. Intrapulmonary expression of CINC gene was assessed by reverse transcription polymerase chain reaction(RT-PCR). Results Compared with that in controls,intrapulmonary expression of CINC gene?wet to weight ratio,and MPO of lungs were increased greatly in twelve hours. The CINC mRNA expression in lungs was significantly correlated with the severity of lung injury,wet to weight ratio,and MPO of lungs. Conclusion Overpexpression of CINC mRNA in the lungs might play an important role in the pathogenesis of ALI in early SAP.

16.
Medical Journal of Chinese People's Liberation Army ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-554489

ABSTRACT

Objective To study the role of Caspase-1 activated cytokines in acute lung injury (ALI) in experimental severe acute pancreatitis (SAP). Methods A rat model of SAP was induced by retrograde infusion of 5% sodium taurocholate into the biliary pancreatic duct. Thirty-two SD rats were randomly divided into four groups: healthy control (HC), SAP 6h, SAP 12h and SAP 18h groups. Serum IL-1? level was measured by ELISA. Intrapulmonary expressions of Caspase-1, IL-1? and IL-18 mRNA were detected by semi-quantitative RT-PCR. Results The serum IL-1? levels were significantly increased after the induction of pancreatitis (P

17.
Chinese Journal of Digestion ; (12)2001.
Article in Chinese | WPRIM | ID: wpr-683380

ABSTRACT

Objective To construct a nucleic acid vaccine expressing H.pylori HpaA and inter- leukin-2 gene and to identify the immunogenicity of the vaccine proteins in vitro and protection in vivo. Methods The H paA gene fragment was amplified by polymerase chain reaction(PCR) from the genomic DNA of the standard H.pylori strain 17874.Mouse interlukin(IL)-2 gene was amplified from pClneo- IL-2.The HpaA and IL-2 were cloned into pUCmT vector.After DNA sequences of the amplified HpaA gene and IL-2 were confirmed,both were cloned into the eukaryotic expression vector pIRES through a serial of enzyme digestion and ligation reactions.The recombinant plasmids were screened by PCR and restriction enzyme digestion.Then,recombinant pIRES-HpsA-IL-2 was transfected to COS-7 cells using Lipofectamine~(TM)2000.The immunogenicity of HpaA and IL-2 protein was detected by SDS- PAGE and Western blot.The recombinant plasmids were transformed to LB5000 and then to final host SL7207.The recombinant strains were passaged repeatedly.The mice were challenged with H.pylori after 4 weeks of inoculation of nucleic acid vaccine.H.pylori infection was detected by rapid urease test.Results The amplified HpaA gene fragment and IL-2 were confirmed by sequence analysis.The eukaryotic expression vector plRES and the pIRES-HpaA-IL-2 construction were confirmed by PCR and restriction digestion.The expressions of HpaA(30 000) and IL-2(14 000)protein by pIRES-HpaA-IL- 2 were detected by Western blot.The in vivo study showed that 75.0% and 58.4% of mice vaccinated by HpaA-IL-2 and HpaA,respectively,were protected anaigst H.pylory infection,which was signifi- cant different in comparison with PBS control (P

18.
Chinese Journal of Pathophysiology ; (12)2000.
Article in Chinese | WPRIM | ID: wpr-521194

ABSTRACT

AIM: To evaluate the dynamic changes of the function and ultrastructure of gastric parietal cells under stress and their relation with acute gastric mucosal lesions. METHODS: Thirty-two male SD rats were randomly divided into four groups, which were control group and 1,2,4 h groups under water restraint stress (WRS). The gastric fluid pH value and gastric mucosal ulcer index(UI) were measured. The ultrastructural changes of parietal cells were observed by transmission electronic microscopy (TEM). RESULTS: The study demonstrated that gastric acid secretion increased and gastric fluid pH value decreased gradually and significantly under WRS compared with control group( P

19.
Academic Journal of Second Military Medical University ; (12)1999.
Article in Chinese | WPRIM | ID: wpr-554551

ABSTRACT

Helicobacter pylori(Hp) infection plays an important role in the development of gastric carcinoma. Colonization of CagA-positive Hp is a great risk for intestinal gastric carcinoma. The mechanism of Hp infection changing the gastric epithelium at the molecular level is complex and through multiple pathways. Animal models can be of great help in exploring the role of Hp infection in gastric cancer development. This paper reviewed the latest studies of epidemiology, animal model and molecular pathogenesis of Hp associated gastric cancer.

20.
Chinese Journal of Digestive Endoscopy ; (12)1996.
Article in Chinese | WPRIM | ID: wpr-521686

ABSTRACT

0. 05 ) . Conclusion The cagG gene of Hp was quite conservative, most of the Hp strains in Chinese patients were cagG positive, it has no specific predisposition to different disorders and shows no significant correlation to the extent of gastric mucosal inflammation. Therefore cagG can' t be served solely as a related pathogenic gene for certain diseases.

SELECTION OF CITATIONS
SEARCH DETAIL