[ Objective ] The aim of the study was to construct associated DNA vaccine of PRRS (Porcine reproductive and respiratory syndrome) and PCV-2 (Porcine circovirus type 2) disease and study its immunogenicity. [ Meth...[ Objective ] The aim of the study was to construct associated DNA vaccine of PRRS (Porcine reproductive and respiratory syndrome) and PCV-2 (Porcine circovirus type 2) disease and study its immunogenicity. [ Method] In_ this study, the ORF5 gene of PRRSV isolated in Liaoning was cloned into plRES-neo expression vector, and the neo gene of plRES-neo expression vector was substituted by the ORF2 gene of the PCV-2 Mongolia strain to construct the recombinant expression vector. The expression in BHK cells was detected through Western blot and IFA. Then the ELISA antibody level and the number of spleen T lymphocytes were detected after Balb/c mice were immunized with this DNA vaccine. E Result] The recombinant plasmid plRES-ORF2-ORF5 was constructed successfully and could express the target proteins in BHK cells, as indicated by Western blot and IFA. There was no significant difference in ELISA antibody between plRES-ORF2-ORF5 immunized group and inactived vaccine immunized groups, while the number of spleen T lymphocytes induced by DNA vaccine was higher than that induced by inactived vaccine. [ Conclusion] The recombinant plasmid plRES-ORF2-ORF5 should induce good humoral immune response and cellular immune response in mice, providing the conditions for better prevention and control of PRRS and PCV-2 disease.展开更多
Cathepsin D (CTSD) is a lysosomal acidic endoproteinase that plays an important role in immune response, In this study, we obtained sweetfish (Plecoglossus altivelis) CTSD (PaCTSD) via de-novo transcriptome sequ...Cathepsin D (CTSD) is a lysosomal acidic endoproteinase that plays an important role in immune response, In this study, we obtained sweetfish (Plecoglossus altivelis) CTSD (PaCTSD) via de-novo transcriptome sequencing of sweetfish macrophages. The full length cDNA sequence of PaCTSD was 1955 bp encoding a propeptide of 397 amino acids. The deduced protein had a calculated molecular weight of 43.17x 103. Multiple alignment with other known CTSD amino acid sequences revealed amino acid conservation through the teleosts. Phylogenetic tree analysis showed that PaCTSD grouped tightly with other fish CTSD, and was close to that of Atlantic salmon and rainbow trout. Subsequently, PaCTSD was prokaryotically expressed and refolded by the urea gradient method on a nickel-nitrilotriacetic acid column. Enzyme activity analysis showed that PaCTSD exhibited pH-dependent proteolytic activity. Quantitative real-time PCR showed that PaCTSD mRNA was expressed in all detected tissues in healthy sweetfish. The highest expression was observed in the spleen and white blood cells, followed by liver, head-kidney, kidney, intestine, gill, and muscle. After Listonella anguillarum infection, PaCTSD transcripts were up-regulated significantly in liver, spleen, white blood cells, and head-kidney of sweetfish. In summary, PaCTSD has proteolytic activity and is closely involved in the immune response of sweetfish.展开更多
Objective: Elf-1 is a member of the proto-oncogenes Ets-related transcription factor family and over-expressed in many human tumors, Ki-67 is an important nuclear antigen associated with cell proliferation. This study...Objective: Elf-1 is a member of the proto-oncogenes Ets-related transcription factor family and over-expressed in many human tumors, Ki-67 is an important nuclear antigen associated with cell proliferation. This study investigated the expression of Elf-1 and Ki-67 in non-small-cell lung cancer(NSCLC) and studied their correlation with the clinicopathological features. Methods: Tissue microarray from 64 cases lung cancer tissue and 10 cases normal lung tissue was constructed, immunohistochemical method was used to evaluate the protein expressions of Elf-1 and Ki-67, correlations of the expression of Elf-1 and Ki-67 to clinicopathological features of NSCLC were analyzed. Results: Expression of Elf-1 and Ki-67 in NSCLC tissues were significantly higher than in normal lung tissues(P < 0.05), the positive rate of Elf-1 and Ki-67 was 73.44% and 64.06% in NSCLC group, Overexpression of Elf-1 in NSCLC was significantly related to histopathological grading, different clinical staging and the intensity of ELF-1 expression was significantly higher in the group with lymph node metastasis than that without(P < 0.05). Overexpression of Ki-67 was also closely related to tumor differentiation, clinical stages and lymph node metastasis(P < 0.05). In addition positive correlation was found between the expressive intensity of Elf-1 and Ki-67(τ = 0.295, P = 0.018). Conclusion: The high expression and positive correlation of Elf-1 and Ki-67 in NSCLC suggest that they probably play a role in onset and progression of lung cancer, united detecting their expression could be used as an valuable molecular biological index for predicting the malignant behavior and early diagnosis of NSCLC.展开更多
Objective: The aim of the study was to analyze the clinicopathologic characteristics of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN) and to explore the prognostic factors for patients and differences of...Objective: The aim of the study was to analyze the clinicopathologic characteristics of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN) and to explore the prognostic factors for patients and differences of immunohisto- chemical markers between neuroendocrine tumor (NET) and neuroendocrine carcinoma (NEC). Methods: Retrospective reviews were conducted for the charts of 119 patients with GEP-NEN at the Affiliated Hospital of Qingdao University (China) from August 2003 to December 2013. Kaplan-Meier method was used to do the overall survivals analysis for the patients at different levels of predictive factors. Meanwhile, Cox proportional hazard model was used to select independent risk factors of surJival. Analysis of variance was used to compare the expression of immunohistochemical markers among different patho- logical grades. Results:Among 119 patients, pancreas (45/119, 37.82%) and rectum (33/119, 27.73%) were mostly involved. The onset age of GEP-NEN in female group was younger than that of the male group. There were 13 deaths (10.92%) during 18.9 (0.1-133.4) months follow-up period. Multivariate analysis indicated that neural invasion, gender and pathological grades of NET and NEC were independent risk factors. In neuroendocrine neoplasm (NEN), Syn expression in G2 was higher than G1 and G3, while CgA showed no significant difference. All markers showed no significant differences between NET and NEC. Conclusion: GEP-NEN may occur at multiple sites of digestive system and lack specific clinical manifestations. Syn expression detected for the prognosis of G1, G2 and G3 tumors have clinical significance. Neural invasion, sex and patho- logical grades were independent prognostic factors for GEP-NEN patients. No significant difference was found in different pathological grades of NET and NEC.展开更多
Objective:Germline mutations in mismatch repair(MMR)genes cause Lynch syndrome(LS).LS is an inherited disease,and an important consequence of MMR deficiency is microsatellite instability(MSI)phenotype.MSI phenotype in...Objective:Germline mutations in mismatch repair(MMR)genes cause Lynch syndrome(LS).LS is an inherited disease,and an important consequence of MMR deficiency is microsatellite instability(MSI)phenotype.MSI phenotype influences the efficacy of5 fluorouracil(5-FU)chemotherapy.Reproducible,cost effective,and easy to perform laboratory tests are required to include MSI detection in routine laboratory practice.Evaluation of CAT25 as monomorphic short tandem repeat sequence enables CAT25 to be an efficient screening tool among hereditary nonpolyposis colorectal cancer(HNPCC)patients compared with other methods used currently.Methods:Based on Amsterdam II criteria,31 patients in 31 families were shortlisted from a total number of 1,659 colorectal cancer patients.MSI status was examined in these patients using CAT25 and a commercially available Promega MSI five-markerbased detection system as well as immunohistochemical(IHC)staining of four important MMR proteins.Patients were scored as high microsatellite instable(MSI-H),low(MSI-L),or stable(MSS).MSI status determined by CAT25 single mononucleotide marker was compared with that of five mononucleotide markers,Promega commercial kit,and IHC method.Results:MMR protein deficiency was observed on 7/31 probands using IHC methodology and 6/31 categorized as MSI-H using commercial kit or CAT25 single marker.The sensitivity and specificity of the CAT25 single marker were the same as those detected by five-marker Promega commercial kit in our patients.Conclusions:Based on our results,the performance of the CAT25 single mononucleotide marker for MSI status determination in our HNPCC patients is the same as that of the five-marker-based commercial kit.展开更多
Irritable bowel syndrome (IBS) is defined by the Rome ]I[ criteria as symptoms of recurrent abdominal pain or discomfort with the onset of a marked change in bowel habits with no evidence of an inflammatory, anatomi...Irritable bowel syndrome (IBS) is defined by the Rome ]I[ criteria as symptoms of recurrent abdominal pain or discomfort with the onset of a marked change in bowel habits with no evidence of an inflammatory, anatomic, metabolic, or neoplastic process. As such, many clinicians regard IBS as a central nervous system problem of altered pain perception. Here, we review the recent literature and discuss the evidence that supports an organic based model, which views IBS as a complex, heterogeneous, inter-dependent, and multi-variable inflammatory process along the neuronal-gut axis. We delineate the organic pathophysiology of IBS, demonstrate the role of inflammation in IBS, review the possible differences between adult and pediatric IBS, discuss the merits of a comprehensive treatment model as taught by the Institute of Functional Medicine, and describe the potential for future research for this syndrome.展开更多
Objective: The aim of the study was to investigate the clinic manifestation, diagnosis and treatment on multiple myeloma (MM) with the onset of renal impairment. Methods: The 27 cases of multiple myeloma with the onse...Objective: The aim of the study was to investigate the clinic manifestation, diagnosis and treatment on multiple myeloma (MM) with the onset of renal impairment. Methods: The 27 cases of multiple myeloma with the onset of renal impair-ment were collected in Department of Nephrology, Wuhan General Hospital of Guangzhou Command, China, from January 2007 to January 2011. All cases were divided into the groups with renal dysfunction (n = 16) and normal renal function (n = 11). The clinic manifestations, treatments and prognosis of all patients were analyzed. Results: Of all the patients in normal renal function group, 5 suffered nephrotic syndrome, 4 had abnormal results of routine urinalysis (hematuria or proteinuria) which were not caused by nephrotic syndrome, and 1 suffered urinary tract infection. Five pathological specimens of renal biopsy revealed that light chain protein, immunoglobulin and complement C3 were deposited mainly in the glomerular base-ment membrane and mesangia, tubular basement membrane and arteriolar walls. Two pathological specimens were proved to be renal amyloidosis. Patients with renal dysfunction had poorer prognosis, severer anemia, higher values of serum lactate dehydrogenase (LDH) and β2-microglobulin (β2-MG), worse responses to chemotherapy. Of 16 patients with renal dysfunc-tion, 14 (87.5%) were stage III, which were significantly higher than that in the group of normal renal function [63.6% (7/11)]. Of 16 cases with renal dysfunction, 9 were treated with blood purification, and 5 of 9 cases were treated with plasma exchange. Conclusion: Multiple myeloma with the onset of renal impairment was easily misdiagnosed. Hemodialysis concomitant with chemotherapy could contribute to recovery of renal function.展开更多
We report a rare case of Pseudo-Meigs' Syndrome caused by ovarian metastasis from sigmoid colon cancer, which was accompanied by peritoneal dissemination. A 58-year-old female patient presented with massive right ...We report a rare case of Pseudo-Meigs' Syndrome caused by ovarian metastasis from sigmoid colon cancer, which was accompanied by peritoneal dissemination. A 58-year-old female patient presented with massive right pleural effusion, ascites and a huge pelvic mass. Under the diagnosis of an advanced ovarian tumor, bilateral oophorectomy was performed and sigmoidectomy was also carried out after intraoperative diagnosis of peritoneal dissemination involving the sigmoid colon. How- ever, immunohistochemical staining revealed that the ovarian lesions were metastasis from the primary advanced colon cancer. Postoperatively, ascites and pleural effusion subsided, and the diagnosis of Pseudo-Meigs' Syndrome due to a metastatic ovarian tumor from colon cancer was determined. The patient is now undergoing a regimen of chemotherapy for colon cancer without recurrence of ascites or hydrothorax 10 mo after the surgery. Pseudo-Meigs' Syndrome due to a metastaticovarian tumor from colon cancer is rare but clinically important because long-term alleviation of symptoms can be achieved by surgical resection. This case report suggests that selected patients, even with peritoneal dissemination, may obtain palliation from surgical resection of metastatic ovarian tumors.展开更多
Forty-six cases of Behcet's disease were randomly divided into two groups. The 26 cases in the treatmentgroup were treated by acupuncture and the 20 cases in the control group with the drugs. The level ofL-chain (...Forty-six cases of Behcet's disease were randomly divided into two groups. The 26 cases in the treatmentgroup were treated by acupuncture and the 20 cases in the control group with the drugs. The level ofL-chain (κ) of IgM and the level of the trace element Zn were determined before and after treatment inthe treatment group. The results showed that the recurrence rate in the treatment group was significantlylower than that in the control group (P<0.01), and the differences in the level of L-chain (κ) of IgM andlevel of Zn in the treatment group before and after treatment were very significant (P<0.01). These twoindexes tended to become normal after treatment.展开更多
文摘[ Objective ] The aim of the study was to construct associated DNA vaccine of PRRS (Porcine reproductive and respiratory syndrome) and PCV-2 (Porcine circovirus type 2) disease and study its immunogenicity. [ Method] In_ this study, the ORF5 gene of PRRSV isolated in Liaoning was cloned into plRES-neo expression vector, and the neo gene of plRES-neo expression vector was substituted by the ORF2 gene of the PCV-2 Mongolia strain to construct the recombinant expression vector. The expression in BHK cells was detected through Western blot and IFA. Then the ELISA antibody level and the number of spleen T lymphocytes were detected after Balb/c mice were immunized with this DNA vaccine. E Result] The recombinant plasmid plRES-ORF2-ORF5 was constructed successfully and could express the target proteins in BHK cells, as indicated by Western blot and IFA. There was no significant difference in ELISA antibody between plRES-ORF2-ORF5 immunized group and inactived vaccine immunized groups, while the number of spleen T lymphocytes induced by DNA vaccine was higher than that induced by inactived vaccine. [ Conclusion] The recombinant plasmid plRES-ORF2-ORF5 should induce good humoral immune response and cellular immune response in mice, providing the conditions for better prevention and control of PRRS and PCV-2 disease.
基金Foundation items: The project was supported by the Program for the National Natural Science Foundation of China (31201970, 31372555), Zhejiang Provincial Natural Science Foundation of China (LZ13C- 190001, LQ13C190002)
文摘Cathepsin D (CTSD) is a lysosomal acidic endoproteinase that plays an important role in immune response, In this study, we obtained sweetfish (Plecoglossus altivelis) CTSD (PaCTSD) via de-novo transcriptome sequencing of sweetfish macrophages. The full length cDNA sequence of PaCTSD was 1955 bp encoding a propeptide of 397 amino acids. The deduced protein had a calculated molecular weight of 43.17x 103. Multiple alignment with other known CTSD amino acid sequences revealed amino acid conservation through the teleosts. Phylogenetic tree analysis showed that PaCTSD grouped tightly with other fish CTSD, and was close to that of Atlantic salmon and rainbow trout. Subsequently, PaCTSD was prokaryotically expressed and refolded by the urea gradient method on a nickel-nitrilotriacetic acid column. Enzyme activity analysis showed that PaCTSD exhibited pH-dependent proteolytic activity. Quantitative real-time PCR showed that PaCTSD mRNA was expressed in all detected tissues in healthy sweetfish. The highest expression was observed in the spleen and white blood cells, followed by liver, head-kidney, kidney, intestine, gill, and muscle. After Listonella anguillarum infection, PaCTSD transcripts were up-regulated significantly in liver, spleen, white blood cells, and head-kidney of sweetfish. In summary, PaCTSD has proteolytic activity and is closely involved in the immune response of sweetfish.
基金Supported by grants from the Science and Technology Development Project of Yantai(No.2008162)Natural Sciences Foundation of Shandong(No.ZR2013HL004)Science and Technology Important Project of Binzhou Medical College(No.BY20bkjzd04)
文摘Objective: Elf-1 is a member of the proto-oncogenes Ets-related transcription factor family and over-expressed in many human tumors, Ki-67 is an important nuclear antigen associated with cell proliferation. This study investigated the expression of Elf-1 and Ki-67 in non-small-cell lung cancer(NSCLC) and studied their correlation with the clinicopathological features. Methods: Tissue microarray from 64 cases lung cancer tissue and 10 cases normal lung tissue was constructed, immunohistochemical method was used to evaluate the protein expressions of Elf-1 and Ki-67, correlations of the expression of Elf-1 and Ki-67 to clinicopathological features of NSCLC were analyzed. Results: Expression of Elf-1 and Ki-67 in NSCLC tissues were significantly higher than in normal lung tissues(P < 0.05), the positive rate of Elf-1 and Ki-67 was 73.44% and 64.06% in NSCLC group, Overexpression of Elf-1 in NSCLC was significantly related to histopathological grading, different clinical staging and the intensity of ELF-1 expression was significantly higher in the group with lymph node metastasis than that without(P < 0.05). Overexpression of Ki-67 was also closely related to tumor differentiation, clinical stages and lymph node metastasis(P < 0.05). In addition positive correlation was found between the expressive intensity of Elf-1 and Ki-67(τ = 0.295, P = 0.018). Conclusion: The high expression and positive correlation of Elf-1 and Ki-67 in NSCLC suggest that they probably play a role in onset and progression of lung cancer, united detecting their expression could be used as an valuable molecular biological index for predicting the malignant behavior and early diagnosis of NSCLC.
文摘Objective: The aim of the study was to analyze the clinicopathologic characteristics of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN) and to explore the prognostic factors for patients and differences of immunohisto- chemical markers between neuroendocrine tumor (NET) and neuroendocrine carcinoma (NEC). Methods: Retrospective reviews were conducted for the charts of 119 patients with GEP-NEN at the Affiliated Hospital of Qingdao University (China) from August 2003 to December 2013. Kaplan-Meier method was used to do the overall survivals analysis for the patients at different levels of predictive factors. Meanwhile, Cox proportional hazard model was used to select independent risk factors of surJival. Analysis of variance was used to compare the expression of immunohistochemical markers among different patho- logical grades. Results:Among 119 patients, pancreas (45/119, 37.82%) and rectum (33/119, 27.73%) were mostly involved. The onset age of GEP-NEN in female group was younger than that of the male group. There were 13 deaths (10.92%) during 18.9 (0.1-133.4) months follow-up period. Multivariate analysis indicated that neural invasion, gender and pathological grades of NET and NEC were independent risk factors. In neuroendocrine neoplasm (NEN), Syn expression in G2 was higher than G1 and G3, while CgA showed no significant difference. All markers showed no significant differences between NET and NEC. Conclusion: GEP-NEN may occur at multiple sites of digestive system and lack specific clinical manifestations. Syn expression detected for the prognosis of G1, G2 and G3 tumors have clinical significance. Neural invasion, sex and patho- logical grades were independent prognostic factors for GEP-NEN patients. No significant difference was found in different pathological grades of NET and NEC.
文摘Objective:Germline mutations in mismatch repair(MMR)genes cause Lynch syndrome(LS).LS is an inherited disease,and an important consequence of MMR deficiency is microsatellite instability(MSI)phenotype.MSI phenotype influences the efficacy of5 fluorouracil(5-FU)chemotherapy.Reproducible,cost effective,and easy to perform laboratory tests are required to include MSI detection in routine laboratory practice.Evaluation of CAT25 as monomorphic short tandem repeat sequence enables CAT25 to be an efficient screening tool among hereditary nonpolyposis colorectal cancer(HNPCC)patients compared with other methods used currently.Methods:Based on Amsterdam II criteria,31 patients in 31 families were shortlisted from a total number of 1,659 colorectal cancer patients.MSI status was examined in these patients using CAT25 and a commercially available Promega MSI five-markerbased detection system as well as immunohistochemical(IHC)staining of four important MMR proteins.Patients were scored as high microsatellite instable(MSI-H),low(MSI-L),or stable(MSS).MSI status determined by CAT25 single mononucleotide marker was compared with that of five mononucleotide markers,Promega commercial kit,and IHC method.Results:MMR protein deficiency was observed on 7/31 probands using IHC methodology and 6/31 categorized as MSI-H using commercial kit or CAT25 single marker.The sensitivity and specificity of the CAT25 single marker were the same as those detected by five-marker Promega commercial kit in our patients.Conclusions:Based on our results,the performance of the CAT25 single mononucleotide marker for MSI status determination in our HNPCC patients is the same as that of the five-marker-based commercial kit.
基金Supported by A Generous Grant from the Riverside Community Health Foundation
文摘Irritable bowel syndrome (IBS) is defined by the Rome ]I[ criteria as symptoms of recurrent abdominal pain or discomfort with the onset of a marked change in bowel habits with no evidence of an inflammatory, anatomic, metabolic, or neoplastic process. As such, many clinicians regard IBS as a central nervous system problem of altered pain perception. Here, we review the recent literature and discuss the evidence that supports an organic based model, which views IBS as a complex, heterogeneous, inter-dependent, and multi-variable inflammatory process along the neuronal-gut axis. We delineate the organic pathophysiology of IBS, demonstrate the role of inflammation in IBS, review the possible differences between adult and pediatric IBS, discuss the merits of a comprehensive treatment model as taught by the Institute of Functional Medicine, and describe the potential for future research for this syndrome.
文摘Objective: The aim of the study was to investigate the clinic manifestation, diagnosis and treatment on multiple myeloma (MM) with the onset of renal impairment. Methods: The 27 cases of multiple myeloma with the onset of renal impair-ment were collected in Department of Nephrology, Wuhan General Hospital of Guangzhou Command, China, from January 2007 to January 2011. All cases were divided into the groups with renal dysfunction (n = 16) and normal renal function (n = 11). The clinic manifestations, treatments and prognosis of all patients were analyzed. Results: Of all the patients in normal renal function group, 5 suffered nephrotic syndrome, 4 had abnormal results of routine urinalysis (hematuria or proteinuria) which were not caused by nephrotic syndrome, and 1 suffered urinary tract infection. Five pathological specimens of renal biopsy revealed that light chain protein, immunoglobulin and complement C3 were deposited mainly in the glomerular base-ment membrane and mesangia, tubular basement membrane and arteriolar walls. Two pathological specimens were proved to be renal amyloidosis. Patients with renal dysfunction had poorer prognosis, severer anemia, higher values of serum lactate dehydrogenase (LDH) and β2-microglobulin (β2-MG), worse responses to chemotherapy. Of 16 patients with renal dysfunc-tion, 14 (87.5%) were stage III, which were significantly higher than that in the group of normal renal function [63.6% (7/11)]. Of 16 cases with renal dysfunction, 9 were treated with blood purification, and 5 of 9 cases were treated with plasma exchange. Conclusion: Multiple myeloma with the onset of renal impairment was easily misdiagnosed. Hemodialysis concomitant with chemotherapy could contribute to recovery of renal function.
文摘We report a rare case of Pseudo-Meigs' Syndrome caused by ovarian metastasis from sigmoid colon cancer, which was accompanied by peritoneal dissemination. A 58-year-old female patient presented with massive right pleural effusion, ascites and a huge pelvic mass. Under the diagnosis of an advanced ovarian tumor, bilateral oophorectomy was performed and sigmoidectomy was also carried out after intraoperative diagnosis of peritoneal dissemination involving the sigmoid colon. How- ever, immunohistochemical staining revealed that the ovarian lesions were metastasis from the primary advanced colon cancer. Postoperatively, ascites and pleural effusion subsided, and the diagnosis of Pseudo-Meigs' Syndrome due to a metastatic ovarian tumor from colon cancer was determined. The patient is now undergoing a regimen of chemotherapy for colon cancer without recurrence of ascites or hydrothorax 10 mo after the surgery. Pseudo-Meigs' Syndrome due to a metastaticovarian tumor from colon cancer is rare but clinically important because long-term alleviation of symptoms can be achieved by surgical resection. This case report suggests that selected patients, even with peritoneal dissemination, may obtain palliation from surgical resection of metastatic ovarian tumors.
文摘Forty-six cases of Behcet's disease were randomly divided into two groups. The 26 cases in the treatmentgroup were treated by acupuncture and the 20 cases in the control group with the drugs. The level ofL-chain (κ) of IgM and the level of the trace element Zn were determined before and after treatment inthe treatment group. The results showed that the recurrence rate in the treatment group was significantlylower than that in the control group (P<0.01), and the differences in the level of L-chain (κ) of IgM andlevel of Zn in the treatment group before and after treatment were very significant (P<0.01). These twoindexes tended to become normal after treatment.