Objective To investigate the clinico-pathological characteristics of nodular goiter (NG) with lymphocytes infiltration and its relationship with Hashimoto′s disease. Methods Clinical and histologic data were collected and reviewed for 1121 cases of nodular goiter and 55 cases of thyroid adenoma, among which 83 cases had lymphocytes infiltration. These 83 cases of NG with lymphocytes infiltration were observed histologically in detail, followed up and assessed for the correlation of thyroid function to histologic findings. Results The overall incidence of lymphocytic infiltration in non-toxic nodular goiter was 7.4% (83/1121). Most of them were women of middle age. All had lymphocytes infiltration, follicule atrophy and oxiphilic epithelia in the background of nodular goiter. Seventy-six percent underwent cystic degeneration or had fibrous scar. Nineteen percent with lymphocytes infiltration in NG developed post-operative hypothyroidism. Six cases (7%) had high levels of thyroid auto-antibodies. None of the 55 cases of thyroid adenoma had overt lymphocytes infiltration. Conclusion Nodular goiter with lymphocytes infiltration and Hashimoto′s disease shares similar morphology and clinical presentation in some degree. It seems that NG with lymphocytes infiltration is a kind of inflammatory hyperplasia relating to autoimmunity and perhaps the beginning of some cases of Hashimoto′s disease.

Objective To explore the operation opportunity of cholecystectomy for senile patients with acute cholecystitis combined with underlying disease.Methods 82 elderly patients with acute cholecystitis complicated with underlying diseases underwent cholecystectomy were selected,they were divided into two groups according to the different operation time:the early operation group and the delayed operation group,41 cases in each group.The patients in early operation group were given operation in 72h,while the delayed operation group was given operation more than 72h after attack.The operation time,intraoperative blood loss,intraoperative tissue adhesions,intraoperative exudation,hospitalization time,cost of hospitalization and complications of the two groups were observed and com-pared.Results The operation time of the two groups were (45.32 ±1 2.23)min,(50.89 ±1 3.34)min,there was no significant difference (P >0.05),the intraoperative tissue adhesions of the delayed operation group was more than the early operation group,and the intraoperative hemorrhage and exudation in the early operation group were significantly higher than those of the delayed operation group (P <0.05),and the hospital stay and cost of hospitalization were sig-nificantly lower than those of the delayed operation group (P <0.05).1 case in the early operation group occurred pulmonary infection,3 cases with incision infection,the incidence of complications was 9.76%;1 case in the delayed operation group underwent laparotomy operation,2 cases in the delayed operation group had pulmonary infection,inci-sion infection in 4 cases,the incidence rate of complication was 1 4.63%,there was no significant difference between the two groups (P >0.05).Conclusion The optimal operation timing of acute cholecystitis combined with basic dis-eases in the elderly patients underwent cholecystectomy is within 72h.For patients who can not be immediately given operation due to various reasons,delayed cholecystectomy is safe and feasible.

The aim of this study is to construct a bifunctional fusion protein, which can conjugate both human red blood cells and antibodies against classical swine fever virus (CSFV). We respectively amplified 2E8ScFv and mE2 genes from different recombinant vectors, in which 2E8ScFv gene is the single chain Fv gene against H antigen of human red blood cells, whereas mE2 gene is the main antigen coding region gene of CSFV E2 protein. We used overlap extension PCR to obtain an artificial fusion gene segment 2E8mE2 containing genes of Both 2E8ScFv and mE2, then ligated into the expression vector pET-DsbA and expressed in Escherichia coli BL21(DE3) PlysS host cells, after induced with IPTG the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. We purified the fusion protein and renatured it from inclusion bodies to obtain a native state of well biological activity. The Erythrocyte agglutination test results indicated that the fusion protein can conjugate both human red blood cells and antibodies of CSFV.
Erythrocytes ; immunology ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunoglobulin Fragments ; biosynthesis ; genetics ; immunology ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Single-Chain Antibodies ; biosynthesis ; genetics ; Viral Envelope Proteins ; biosynthesis ; genetics

OBJECTIVETo develop monoclonal antibodies against the catalytic subunit of human telomerase hTERT for its expression detection of human tumors.

METHODSA dominant epitope in hTERT (peptide hTERT(9))was automatically synthesized based on Fmoc method, and was used to immunize BALB/c mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, and the characterization of which were performed by Western blotting and immunohistochemical staining.

RESULTSAntigenic peptide hTERT(9) was synthesized and confirmed by MALDI-TOF-MS and HPLC analysis. Three hybridoma cell lines secreting anti-hTERT(9) antibodies designated as H4, G8 and A11 were established after primary screening and consequent three rounds of limited dilution. Both of H4 and G8 were IgM, while A11 was IgG1 in isotyping. The competitive assay showed that the antibodies were hTERT(9) specific, and the affinity of G8 was stronger than that of H4 and A11 assayed by affinity ranking. However, in Western blotting, both of H4 and G8 stained an about 123 000 protein band with HeLa and 293 cell extracts but not with normal 2BS cells. Besides, positive staining presented in the nucleus of HeLa, while 2BS was non-reactive immunohistochemically. The sections from paraffin-embedded blocks of 127 cases of human cancer, 40 of precancerous and 19 of benign tumors were in situ stained by G8 antibody, the results showed that the human cancer tissues were 80.31% (102/127) positive in specific nuclear reaction, on the contrary, only a minority of precancerous lesions present weak positive (17.5%, 7/40), and negative in benign tumors (0/19).

CONCLUSIONSThe monoclonal antibodies developed against synthetic peptide were hTERT-specific and could recognize both the native and the denatured form. Thus their use in immunoblotting or immunohistochemistry for detecting the telomerase hTERT expression of cancer cell and tissues was promising.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Binding, Competitive ; Blotting, Western ; methods ; Catalytic Domain ; DNA-Binding Proteins ; Female ; HeLa Cells ; Humans ; Immunohistochemistry ; methods ; Mice ; Mice, Inbred BALB C ; Neoplasms ; enzymology ; pathology ; Telomerase ; chemical synthesis ; immunology