OBJECTIVETo investigate the effects of small interfering RNA targeting connective tissue growth factor (CTGF) on rat transforming growth factor beta (TGF beta)/Smads signal pathway.

METHODSChemically synthetic siRNA targeting CTGF was transfected into HSC T6 and then they were injected into rat livers through their intraportal veins. At the same time these rats also received CCl4 subcutaneously every three days for 6 consecutive weeks. Untreated HSC T6 or/and rats with random siRNA treatment served as controls. Total RNA or/and protein in HSC T6 and rat hepatic tissues were extracted. The expressions of CTGF and TGF beta 1, Smad2, 3 and 7 genes were detected by reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot.

RESULTSCTGF siRNA significantly reduced the expression of CTGF protein in HSC T6. At 48 h after CTGF siRNA treatment, the down-regulation of CTGF protein was the most significant, up to 94%+/-4% (t=46.196, P less than 0.01), but the expressions of TGF beta 1, Smad2, 3 and 7 mRNA showed no differences in HSC T6 compared with the blank controls. Six weeks after CCl4 injections, prominent up-regulations were observed in the gene expressions of CTGF and TGF beta 1 in saline control or siRNA-treated rat livers. Administering CTGF siRNA for six weeks markedly attenuated the induction of CTGF and TGF beta 1 genes; the expressions of CTGF and TGF beta 1 protein decreased by 95%+/-2% (F=21.234, P less than 0.01) and 74%+/-8% (F=13.464, P less than 0.05), respectively, whereas Smad2, 7 protein expressions were not affected.

CONCLUSIONSilencing the CTGF gene can suppress the TGF beta /Smads signal pathway in rat livers.

Animals ; Connective Tissue Growth Factor ; metabolism ; Gene Silencing ; Male ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Smad Proteins ; metabolism ; Transfection ; Transforming Growth Factor beta ; metabolism

Objective To analyze the outcome of surveillance results on plague and to provide the evidences for the policy making in Longlin county Guangxi. Methods The epidemic data and the surveillance results of plague were analyzed and assessed with epidemiology methods in Longlin county Guangxi from 2000 to 2009, and the density of rodents, the rodents infected with flea, flea index and other indicators were calculated. Regional composition of the rats and fleas were analyzed. Results A totally of 4829 rats were captured and 4737 fleas were collected in the past 10 years, Rattus Flavipestus(81.92%,3956/4829) and Xenopsylla Cheopis (79.04%,3744/4737) were dominant species. The annual average density of rodents, the rodents infected with flea, index of flea were 3.30%(4829/146 206), 27.99%(1351/4827) and 0.98(4737/4827), respectively. A totally of 4792 rats were examined and 10 strains Yersinia Pestis were isolated. Indirect hemorrhagic assessed(IHA) was used to test the F1 antibody against plague in the blood serum of the rats and indicator animals, and 3 positive rats and 24 positive animals were found, respectively. Twenty seven natural villages in 3 towns had been involved in the plague. Conclusions The plague foci exists in Longlin county of Guangxi province. The plague foci in the areas have the same feature with the plague foci of Rattus Flavipectus. There is a potential risk for plague in this region, we should improve the quality of surveillance, increase indicator animals of the plague, and try to apply new surveillance method.

Objective To evaluate contrast-enhanced fluid-attenuated inversion recovery (FLAIR) imaging in the detection of leptomeningeal lesions.Methods Seventeen patients with a variety of leptomeningeal lesions were analyzed.The MRI protocol included un-enhanced and contrast-enhanced FLAIR images and contrast-enhanced T_1WI,Comparisons between contrast-enhanced FLAIR images and T_1WI and between un-enhanced and contrast-enhanced FLAIR images were made to determine which sequence better depicted the lesions.Results Leptomeningeal lesions showed as either diffusely or locally abnormal hyper-intensity along sulci or cistern on three sequences.Comparison between contrast-enhanced FLAIR and T_1WI showed that only contrast-enhanced FLAIR revealed the abnormalities in 7,both revealed the abnormalities but the former was superior in 2 ,and both were conspicuous in 7. In 1 patient of tuberculous meningitis,diffuse abnormalities of sulci were shown only on contrast-enhanced FLAIR, abnormalities of cisterns were shown on both sequences but the former was superior.Comparison between un- enhanced and contrast-enhanced FLAIR showed that only contrast-enhanced FLAIR revealed the abnormalities in 9,both revealed the abnormalities but the former was superior in 3,and both were conspicuous in 4. In 1 patient of tuberculous meningitis,abnormalities of cisterns were shown only on contrast-enhanced FLAIR,diffuseabnormalities of sulci were shown on both sequences but the former was superior.Conclusions Contrast-enhanced FLAIR images were superior to un-enhanced FLAIR images and contrast-enhanced T_1WI in the detection of leptomeningeal lesions. Contrast-enhanced FLAIR images are helpful and should be considered when findings on un-enhanced FLAIR images and/or contrast-enhanced T,WI are inconclusive.

Objective:to explore the effect of nano-carbon tracer on the dissection of central group lymph nodes in thyroid cancers.Methods:60 patients with thyroid cancers enrolled from January 2015 to December 2015 in our hospital were selected as research objects.Tracing group contained 30 cases would carry out nano-carbon tracer for the dissection of lymph nodes,while the other 30 patients without using nano-carbon tracer were defined as control group.The number of dissected lymph nodes,the discovery rate of positive lymph nodes and the postoperative parathyroid function were made a comparison between the two groups.Results:the total number of dissected lymph nodes in the tracing group was more than the control group (269 vs 204).The average number of dissected lymph nodes in the tracing group (8.97 ± 1.65/case) was also significantly more than the control group(6.8 ± 1.52/case)(P＜0.05).In the tracing group,the total discovery rate of positive lymph nodes was 40.15％,while the control group was 37.25％.Therefore,the average number of dissected positive lymph nodes in the tracing group (3.6 ± 1.16/case) was significantly more than the control group (2.53 ± 1.17/case)(P＜0.05).Observation of the postoperative adverse reactions,there were fewer patients suffering hypocalcemia or recurrent laryngeal nerve injury in the tracing group compared to the control group.In detail,although the blood calcium levels on the 2nd day after operation in both two groups decreased compared with preoperative baseline values,significantly statistical difference was only observed in the control group with 2.173 ±0.20mmol/L in postoperation vs 2.28 ± 0.06mmol/L in pre-operation (P＜0.05).What's more,the blood calcium level in the tracing group on the 2nd day after operation (2.27 ± 0.19mmol/L) was significantly higher than the control group (2.173 ± 0.20mmol/L)(P＜0.05).Besides,the postoperative PTH levels in both two groups reduced in some degree compared to the preoperative baseline values,but there were no statistical differences (P＞0.05).Conclusion:using nano-carbon tracer during the operation would be benefit for the dissection of positive central group lymph nodes,the recognition of parathyroid glands and reduction of postoperative adverse reactions.

Catheterization ; adverse effects ; Child ; Humans ; Jugular Veins ; injuries ; Male ; Punctures

OBJECTIVETo observe the morphology and phenotypes of cells extracted from the endplate in the intervertebral discs and identify the factors affecting their biological characteristic.

METHODSThe intervertebral disc endplate were digested enzymatically, and the morphology of the obtained cells was examined under light microscope. Immunhistochemical analysis of collagen II and real-time PCR was carried out, and the morphologies, viability, cell growth, apoptosis and chondrocyte matrix production were compared between the cells isolated from the degenerative and normal vertebral endplates.

RESULTSThe cells in primary culture presented with spherical and oval morphology, and the cytoplasm was stained blue with toluidine blue. The morphologies of the cartilage endplate cells and the articular cells were almost identical. All the freshly isolated cells expressed collagen II. The degenerative vertebral endplate cells showed decreased expression of collagen II with increased apoptotic cells as compared with normal vertebral endplate cells.

CONCLUSIONThe intervertebral disc endplate cells, like articular cartilage cells, express cartilage-specific matrix proteins. Degenerative vertebral endplate cells show decreased cell vitality with increases cell apoptosis.

Adult ; Apoptosis ; physiology ; Cartilage ; metabolism ; pathology ; Cells, Cultured ; Chondrocytes ; metabolism ; pathology ; Collagen ; metabolism ; Female ; Growth Plate ; metabolism ; pathology ; Humans ; Intervertebral Disc ; metabolism ; pathology ; Intervertebral Disc Degeneration ; metabolism ; pathology ; Lumbar Vertebrae ; metabolism ; pathology ; Male ; Young Adult

OBJECTIVETo study the effect of zoledronic acid on cell cycle blocking and induction of apoptosis in lung cancer cell line 95D cells, and their mechanisms of action.

METHODSThe effect of zoledronic acid (ZOL) on proliferation of lung cancer cell line 95D cells was observed by MTT assay. Cell cycle and apoptosis of the lung cancer cells was examined by flow cytometry. The apoptosis in the cancer cells was also examined by light and transmission electron microscopy. The expressions of ERK, Bcl-2, Bax and survivin were measured by Western blot and RT-PCR.

RESULTSZOL showed inhibitory effect on the proliferation of lung cancer cells in vitro, in a time-dependant and a dose-dependant manner. With time extending after ZOL treatment, the number of apoptosis cells was increased. The expression of ERK, Bcl-2 and survivin was down-regulated and that of Bax up-regulated.

CONCLUSIONZoledronic acid can block the cell cycle and induce apoptosis in lung cancer cells in vitro.

Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Diphosphonates ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Imidazoles ; administration & dosage ; pharmacology ; Inhibitor of Apoptosis Proteins ; Lung Neoplasms ; metabolism ; pathology ; Microtubule-Associated Proteins ; genetics ; metabolism ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

Systemic lupus erythematosus (SLE) and clear cell renal cell carcinoma (CC-RCC) are serious disorders and usually fatal, and always accompanied with pathological changes in the kidney. Signal-induced proliferation-associated protein 1 (SIPA-1) is a Rap1GTPase activating protein (Rap1GAP) expressed in the normal distal and collecting tubules of the murine kidney. Lupus-like autoimmune disease and leukemia have been observed in SIPA-1 deficient mice, suggesting a pathological relevance of SIPA-1 to SLE and carcinoma in human being. The expression pattern of SIPA-1 is as yet undefined and the pathogenesis of these diseases in humans remains elusive. In this study, we used both immunohistochemistry and quantum dot (QD)-based immunofluorescence staining to investigate the expression of SIPA-1 in renal specimens from SLE and CC-RCC patients. MTT assay and Western blotting were employed to evaluate the effects of SIPA-1 overexpression on the proliferation and apoptosis of renal cell lines. Semi-quantitative reverse transcriptase-PCR (RT-PCR) was applied to examine the changes of hypoxia-inducible factor-1α (HIF-1α) mRNA level. Results showed that SIPA-1 was highly expressed in the proximal and collecting tubules of nephrons in SLE patients compared to normal ones, and similar results were obtained in the specimens of CC-RCC patients. Although SIPA-1 overexpression did not affect cellular proliferation and apoptosis of both human 786-O renal cell carcinoma cells and rat NRK-52E renal epithelial cell lines, RT-PCR results showed that HIF-1α mRNA level was down-regulated by SIPA-1 overexpression in 786-O cells. These findings suggest that SIPA-1 may play critical roles in the pathological changes in kidney, and might provide a new biomarker to aid in the diagnosis of SLE and CC-RCC.
Apoptosis ; Base Sequence ; Cell Line ; Cell Proliferation ; DNA Primers ; GTPase-Activating Proteins ; metabolism ; Humans ; Kidney Tubules, Proximal ; metabolism ; pathology ; Lupus Erythematosus, Systemic ; metabolism ; pathology ; Nuclear Proteins ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the reliability and feasibility of sentinel lymph node biopsy (SLNB) of papillary thyroid carcinoma using combination of lymphoscintigraphy, the gamma probe and methylene blue staining techniques.

METHODSTwenty-three consecutive patients with thyroid papillary carcinoma were entered in the study between August 2007 and August 2009. All cases were without clinical evidence of cervical lymph node involvement. The (99)Tc(m)-dextran of 74 MBq (2mCi) was injected intratumorally under ultrasound guidance about 2 h to 5 h prior to surgery. Methylene blue was injected around the tumor during surgery. Preoperative lymphoscintigraphy, intra-operative hand-held gamma probe detecting and methylene blue staining techniques were used to detect the sentinel lymph node (SLN). SLN biopsies were sent to prepare frozen sections and the results were compared with specimen of routine selective neck dissection.

RESULTSThe SLNs were identified in all cases with the combination techniques. The SLN identification rates were 87.0% and 100% with methylene blue staining and lymphoscintigraphy plus probe scanning, respectively. Metastases in SLNs were revealed by frozen-section histology in 12 patients. In one case, SLNs frozen-section were negative, but metastasis was detected in routine histology. In other case both SLN and routine histology were negative, but metastasis was detected in non-SLN (level VI) neck dissection. The overall accuracy of the SLN biopsy was 91.3%, positive predictive value 100% and negative predictive value 81.8%.

CONCLUSIONThe results seem the SLN biopsy technique is a feasible and valuable method for detecting cervical lymph node metastasis and is helpful to decide performing neck dissection in patients with cN0 papillary thyroid carcinoma.

Adult ; Aged ; Carcinoma, Papillary ; diagnostic imaging ; pathology ; surgery ; Dextrans ; Female ; Humans ; Lymph Node Excision ; Lymph Nodes ; diagnostic imaging ; pathology ; Lymphatic Metastasis ; Male ; Methylene Blue ; Middle Aged ; Neoplasm Staging ; Organotechnetium Compounds ; Radionuclide Imaging ; Sentinel Lymph Node Biopsy ; Thyroid Neoplasms ; diagnostic imaging ; pathology ; surgery ; Young Adult

OBJECTIVETo study the impact of an agonist anti-CD(40) monoclonal antibody 5C11 on the induction and biological characteristics of leukemic dendritic cells.

METHODSCombinations of 5C11 and different cytokines were used to induce differentiation of leukemic blasts into dendritic cells. Morphology was observed by light microscopy. Surface antigens of the induced cells were analyzed by fluorescence-activated cell sorting (FACS), the yields of dendritic cell by cell counting, the levels of IL-6 and IL-12 by ELISA, T cell proliferating activity by allo-mixed lymphocyte reaction (MLR) in vitro. Allogeneic T cells were stimulated with leukemic dendritic cells and T-cell cytotoxicity was measured by MTT assay.

RESULTSWhen cultured with combinations of 5C11 and different cytokines, the leukemic cells isolated from the patients could differentiate into dendritic cells. The morphology showed typical features of dendritic cells, which expressed high levels of CD(40), CD(80) and CD(86). In comparison with the original leukemia cells, the leukemic dendritic cells secreted less IL-6 but more IL-12 (P < 0.05). The leukemic dendritic cells were potent to stimulate the proliferation of allogeneic T cells, and the latter was able to lyse the original leukemia cells.

CONCLUSIONLeukemic blasts could be induced to differentiate into functional dendritic cells. It may be of great value in the adoptive immunologic therapy of leukemia.

Antibodies, Monoclonal ; immunology ; CD40 Antigens ; physiology ; Cell Differentiation ; Dendritic Cells ; immunology ; Humans ; Immunophenotyping ; Immunotherapy ; Interleukin-12 ; biosynthesis ; Interleukin-6 ; biosynthesis ; Leukemia ; immunology ; pathology ; therapy