Effect of teriparatide on osteoblast differentiation in high glucose environment through Wnt3a / β-catenin pathway / 安徽医科大学学报

Objective @#To investigate the effect of teriparatide ( TPTD) on the generation of MC3T3-E1 cells to- wards osteogenic differentiation via the Wnt3a / β-catenin pathway in a high-glucose environment．@*Methods @#The experiment was divided into five groups : low glucose group，low glucose + TPTD group，high glucose group，high glucose + TPTD group，high glucose + TPTD + Wnt3a inhibitor G244-LM group．Cell proliferation activity was de- tected by Calcein-AM and CCK-8 assay，cell mineralized nodule formation was observed by ALP and alizarin red staining，and actin formation was analyzed by immunofluorescence assay. Real-time PCR was performed to detect Wnt3a，β-catenin，Tcf1，OPG and COL Ⅰ mRNA expression. @*Results @#TPTD had no significant effect on the pro- liferative activity of MC3T3-E1 cells under high glucose condition．The ALP staining area，protein activity and aliza- rin red staining area of the cells in the low glucose + TPTD group were higher than those in the other four groups (P ＜0. 05) ; the high glucose group was lower than the low glucose group (P ＜0. 05 ) ; the high glucose + TPTD group was higher than the high glucose group and the high glucose + TPTD + G244-LM group (P＜0. 05) ．The cy- toskeleton in the low glucose + TPTD group was the clearest ; the cytoskeleton was less clear in both the high glucose and high glucose + TPTD + G244-LM groups than in the high glucose + TPTD group．Genes such as Wnt3a，β-cate- nin，Tcf1，OPG and COL Ⅰ had the highest mRNA levels in the cells of the low glucose + TPTD group (P < 0. 05) ; the mRNA levels of all genes were higher in the low glucose group than thosein the high glucose group (P ＜0. 05) ; the mRNA levels of all genes in the cells of the high glucose + TPTD group were higher than those in the high glucose group and the high glucose + TPTD + G244-LM group ( P＜0. 05) ．@*Conclusion @#High glucose inhibi- ted osteoblast differentiation，and TPTD promoted osteoblast differentiation in high glucose environment by regula- ting Wnt3a / β-catenin pathway.

Analysis of the Relationship Between the Quality of Small Biopsy Specimens of Non-small Cell Lung Cancer and the Mutation Rate of EGFR Gene / 中国肺癌杂志

BACKGROUND@#Epidermal growth factor receptor (EGFR) is the gene with the highest mutation rate in non-small cell lung cancer (NSCLC) patients, and the accurate evaluation of its mutational status can facilitate patients receiving targeted drug therapy and thereby prolong patients' survival. The gene testing platform has adequacy requirements for the specimen quality in order to obtain accurate examination results. It has been reported that the number and proportion of tumor cells in samples will affect the detection rate of EGFR gene mutation. The present study aims to analyze the relationship between the quality of small biopsy specimens of NSCLC and the mutation rate of EGFR gene with amplification refractory mutation system (ARSM) test.@*METHODS@#After collecting the clinical characteristics of 299 cases small biopsy of lung adenocarcinoma, DNA concentration of the specimens and the mutational status of EGFR gene, the number and proportion of tumor cells in HE stained sections evaluated using light microscopy, the relationship between specimen quality and the mutation rate of EGFR gene were analyzed.@*RESULTS@#The mutation rates of EGFR for the groups with tumor cell number ≤500 and >500 were 40.7% (11/27) and 43.8% (119/272) respectively, without significant difference (P=0.764). The mutation rates for the groups with DNA concentration ≤20.4 ng/μL and >20.4 ng/μL were 42.7% (64/150) and 44.3% (66/149) respectively, without significant difference (P=0.776). The mutation rates for the groups with tumor cells proportion ≤30% and >30% were 29.4% (20/68) and 47.6% (110/231) respectively, demonstrating significant difference (P=0.008). Multivariate Logistic analysis showed that male, thyroid transcription factor-1 (TTF-1) negative, smoking history and tumor cell proportion less than 30% were main factors that contributes to the low detection rate of EGFR gene mutation.@*CONCLUSIONS@#After meeting the minimum requirements for detection, the EGFR mutation rate is affected by the proportion of tumor cells in the sample. Therefore, it is necessary to re-evaluate the tumor cell proportion in the last section after the genetic test section. For samples with lower tumor cell proportion, enriching tumor cells through microdissection and other methods is recommended for a more accurate detection result. For specimens that cannot be enriched with tumor cells, circulating tumor DNA (ctDNA) test can be performed as a supplement. If the result is still negative, another biopsy should be considered to obtain enough tumor specimens for molecular testing.

Zoledronate regulates osteoclast differentiation and bone resorption in high glucose through p38 MAPK pathway / 南方医科大学学报

OBJECTIVE@#To investigate the effect of zoledronate (ZOL) on osteoclast differentiation and bone resorption under high glucose, and the regulation mechanism of p38 mitogen activated kinase (p38 MAPK) signaling pathway in this process.@*METHODS@#RAW264.7 cells were divided into four groups: low group, high group, low+ZOL group and high+ZOL group after induced into osteoclasts. Cell proliferation activity was determined by MTT assay. The migration of RAW264.7 cells were examined Optical microscopy. Immunofluorescence microscopy was used to observe the cytoskeleton and sealing zones of osteoclasts. After adding group 5: high + ZOL + SB203580 group, trap staining was used to identify the number of positive osteoclasts in each group. The number and area of resorption lacunae were observed by SEM. The mRNA and protein expression of osteoclast related factors were detected by real-time PCR and Western blotting.@*RESULTS@#The cells in the 5 groups showed similar proliferative activity. High glucose promoted the migration of RAW264.7 cells (@*CONCLUSIONS@#High glucose inhibits osteoclast differentiation and bone resorption. ZOL inhibits osteoclast differentiation and bone resorption in high-glucose conditions by regulating p38 MAPK pathway, which can be a new pathway for ZOL to regulate diabetic osteoporosis.

Clinical analysis of 30 cases of Castleman disease with different types of thoracic involvement / 中华血液学杂志

Objective@#To improve the clinical understanding of Castleman disease (CD) with different types of thoracic involvement, including their clinical features, radiological and pathological findings, diagnosis and current treatment strategies.@*Methods@#Retrospective analysis of 30 patients diagnosed with CD with thoracic involvement and hospitalized between June 2009 and May 2019 in The First Affiliated Hospital of Guangzhou Medical University was performed. Patients were divided into three groups for subsequent analysis based on the clinical data: CD with bronchiolitis obliterans (BO) , unicentric Castleman disease (UCD) without BO, and multicentric Castleman disease (MCD) without BO.@*Results@#Among the 30 patients, there were 5 (16.7%) patients diagnosed with BO, 18 (60.0%) patients had UCD without BO and 7 (23.3%) patients had MCD without BO. The average age of MCD without BO patients was significantly older than that of BO and UCD without BO patients[ (49.29±5.39) ys vs (27.20±3.76) ys and (37.17±2.87) ys; P=0.005 and 0.034, respectively) ]. Pulmonary symptoms were commonly seen in BO group (100%) and MCD without BO group (71.4%) . while no pulmonary symptoms were seen in UCD without BO group. Key abnormal laboratory findings were erythrocyte sedimentation rate (ESR) increase (40%in BO group and 57.1% in MCD without BO group) and hypoxia (60% in BO group and 28.6% in MCD without BO group) . Other abnormal laboratory findings seen in MCD without BO group included anemia and IgG increase (both 57.1%) . Notably, all patients in BO group had extremely severe mixed ventilation dysfunction in the lung function test. CT scan showed lung parenchyma involvement in BO group (100%) , in UCD without BO group (11.1%) featured by solitary pulmonary nodule and in MCD without BO group (57.1%) featured by diffuse lesions in bilateral lungs. The size of lymph nodes was significantly smaller in MCD without BO group comparing to that in BO group and UCD without BO group[short diameter (1.83±0.51) cm vs (4.73±1.63) cm and (3.62±0.26) cm; P=0.006 and 0.011, respectively]. All patients (100%) in the BO group had a pathological type of transparent vascular variant while the same pathological type accounts for 88.9% in UCD without BO patients. The predominantly pathological type (57.1%) was plasma cell variant in the MCD without BO group. Oral ulcers presented in all patients in BO group but were relieved after the mass resection and immunomodulatory therapy, but the pulmonary symptoms were still progressively aggravated. Thoracoscopic mass excision was the main treatment for UCD without BO patients while chemotherapy, immunomodulatory and targeted therapy were commonly used for MCD without BO treatment.@*Conclusion@#The age, clinical symptom, laboratory finding, lung function, imaging manifestation, pathology, treatment and prognosis were different among the three groups. This classification could improve clinical understanding of the disease.

Investigation on the Possibility of EGFR Mutation Testing on DNA Extracted from Fixation Liquid of Lung Cancer Biopsy / 中国肺癌杂志

BACKGROUND@#Epidermal growth factor receptor (EGFR) mutation is the most common gene mutation in patients with non-small cell lung cancer (NSCLC). Many international guidelines are recommended to detected the EGFR mutation before the treatment of advanced non-small cell lung cancer. To investigate the possibility of EGFR mutation testing on DNA extracted from fixation liquid of lung cancer biopsy.@*METHODS@#Fixation liquid of lung cancer biopsy was collected and stored at -80 oC after centrifugal. DNA was extracted and EGFR gene mutation was detected by ARMS. Compared with EGFR mutation status of paraffin-embedded tissues, the consistency, the sensitivity and specificity of EGFR mutation testing were analyzed.@*RESULTS@#Among the 28 cases of EGFR mutation positive and 20 cases of EGFR mutation negative previously tested on paraffin-embedded tissue by clinic test, 20 cases with EGFR mutation positive and 20 cases with negative were detected by matched fixation liquid of lung cancer biopsy, respectively. The sensitivity and specificity were 71.4% and 100%. Moreover, 52 paraffin-embedded tissues and matched fixation liquid of lung cancer biopsy with unknown EGFR mutation status were detected, and the EGFR mutation positive rate were 36.5% and 28.8% respectively. The sensitivity and specificity of fixation liquid of lung cancer biopsy were 78.9% and 100.0%.@*CONCLUSIONS@#Extracting the DNA from fixation liquid of lung cancer biopsy may be a kind of feasible way to detect EGFR mutation.

Lung transplantation in treatment of secondary pleuroparenchymal fibroelastosis: report of one case and literature review / 器官移植

Objective To explore the treatment strategies of pleuroparenchymal fibroelastosis (PPFE). Methods A 22-year-old male patient was complicated with PPFE after receiving chemotherapy in combination with stem cell transplantation for lymphoma. He underwent thoracoscopic left lung tongue wedge resection, bilateral pleurodesis followed by allogeneic left lung transplantation. Literature review was performed to analyze the etiology, pathogenesis, imaging features, pathological features and treatment of PPFE. Results The PPFE patient required the non-invasive ventilator for 24 h before lung transplantation. After lung transplantation, the shortness of breath and respiratory failure were cured and the quality of life was significantly improved. No eligible studies was found in the domestic database, and 26 literatures published in English were found in the international databases. Among them, 9 literatures (case reports) were finally included after screening. PPFE could be divided into the primary and secondary categories according to the etiology. The clinical manifestations of PPFE mainly included dry cough, dyspnea on exertion, chest pain, repeated pneumothorax and body weight loss. Chest CT scan demonstrated irregular thickening of the pleura in bilateral upper lungs. Pathological manifestations consisted of evident thickening of the visceral pleura, fibroelastosis and arrangement disorder in the pleura and the underlying pulmonary interstitium. PPFE could progress rapidly. Adrenocortical hormone and other immunosuppressive agents yielded low clinical efficacy and poor clinical prognosis. Lung transplantation was a necessary treatment for PPFE. Conclusions PPFE cannot be effectively treated by conservative therapy. It is recommended to deliver lung transplantation as early as possible.

The Prognostic Significance of Notch1 and Fatty Acid Binding Protein 7 (FABP7) Expression in Resected Tracheobronchial Adenoid Cystic Carcinoma: A Multicenter Retrospective Study / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Adenoid cystic carcinoma (ACC) of the trachea and bronchus is a rare tumor. Although MYB-NFIB oncogene fusion and Notch1 mutation have been identified in ACC, little is known about the expression and clinical significance of Notch1 and its target gene fatty acid binding protein 7 (FABP7) in tracheobronchial ACC. MATERIALS AND METHODS: Primary tracheobronchial ACC that were resected between 1998 and 2014 were identified through the pathology and oncology database from five thoracic oncology centers in China. A tissue array was constructed from the patients’ samples and the expressions of Notch1 and FABP7 were evaluated by immunohistochemistry. The association between the expression of both markers and survival was determined. RESULTS: Overexpression of Notch1 and FABP7, detected in 37.8% and 38.3% of 368 patients with tracheobronchial ACC, respectively, was an independent prognostic indicator for recurrencefree survival (RFS) by multivariable Cox proportional hazard model (p=0.032 and p=0.048, respectively). Overexpression of Notch1, but not of FABP7, predicted overall survival (OS) (p=0.018). When categorized into four groups according to coexpression of Notch1 and FABP7, patients with overexpression of both Notch1 and FABP7 belonged to the group with the shortest RFS and OS (p=0.01 and p=0.048, respectively). CONCLUSION: Expression of Notch1 and FABP7, and coexpression of Notch1 and FABP7, is strongly associated with poor survival in resected tracheobronchial ACC. These data are consistent with the hypothesis that poor differentiation of tracheobronchial ACC correlates with the activation of Notch signaling.

The effect of subcellular localization of tumor BRCA1 on the sensitivity of cell radiation and PARP inhibitor / 临床与实验病理学杂志

Purpose To investigate the effect of subcellular location of tumor BRCA1 on the sensitivity to ionizing radiation (IR) and PARP inhibitor.Methods siRNA of BRCA1 were first used to inhibit endougenous BRCA1 expression in MCF7 cells.Then,plasmids of pCMV-3xFlag-WT-BRCA1,pCMV-3xFlag-NES-BRCA1 and pCMV-3xFlag-NLS-BRCA1 were transfected in MCF7 cells.Immunofluorescence staining was used to detect BRCA1 subcellular location as well as the formation of Rad51 and γ-H2AX foci.Apoptotic cells were analyzed by flow cytometry,and colony formation assay was performed to evaluate the survival of cells.Results There were 47％ cells with nuclear BRCA1,23％ cells with cytoplasmic BRCA1 and 30％ cell with mixed nuclear and cytoplasmic BRCA1 expression in WT-BRCA1 transfected cell.There were 87％ cells with nuclear BRCA1 in NES-BRCA1 transfected cell,and 82％ cells with cytoplasmic BRCA1 in NLS-BRCA1 transfected cell.There were 87％,84％ and 13％ Rad51 foci positive cells at 2 hours after 4 Gy radiation treatment and 22％,25％ and 59％ γ-H2AX foci positive cells at 24 hours after 4Gy radiation treatment in WT-BRCA1,NES-BRCA1 mutant and NLS-BRCA1 mutant transfected cell respectively.ABT-888 and radiation treatment induced more apoptosis and fewer colonies in NLS-BRCA1 transfected cell than WT-BRCA1,NES-BRCA1 mutant transfected cell.Conclusion Subcellular location of BRCA1 might affect homologous recombination repair of DNA double strand breaks and can be used to predict sensitivity to IR and PARP inhibitor.

Detection of cyclohexanone in workplace air by capillary column gas chromatography / 中国职业医学

OBJECTIVE: To establish a methodology for the detection of cyclohexanone in workplace air by capillary column gas chromatography.METHODS: The air sample in the workplace was collected with activated carbon tube and desorbed with carbon disulfide.The extracts were separated by FFAP capillary column and detected with flame ionization detector,and quantified using standard curve.RESULTS: The linearity range of cyclohexanone was 10.00-1 000.00 mg/L,and the correlation coefficient was 0.999 9.The detection limit and the lower limit of quantification were 1.31 and 4.37 mg/L,respectively.The minimum detectable concentration and the minimum quantitative mass concentration were 4.37 and13.11 mg/m~3,respectively.The average desorption efficiency of cyclohexanone was 95.80%-97.88%.The within-run and between-run relative standard deviation were 0.52%-3.12% and 3.28%-4.75% respectively.The samples could be stored at room temperature for 7 days.CONCLUSION: This method is simple and accurate.It is suitable for the determination of a large number of samples.

Targeted exogenous EGFP gene editing in caprine fetus fibroblasts by zinc-finger nucleases / 生物工程学报

Gene knockout by ZFNs (zinc-finger nucleases) is efficient and specific, and successfully applied in more than 10 organisms. Currently, it is unclear whether this technology can be used for knocking-out enhanced green fluorescent protein (EGFP) gene in transgenic goats. Here we constructed and used ZFN-coding plasmids to produce genetic knockouts in the cells of cloned fetus produced from donor cells by microinjection of EGFP gene. Following introduced plasmids into caprine primary cultured fetus fibroblasts by electroporation, targeting of a transgene resulted in sequence mutation. Using the flow cytometric analysis, we confirmed the disappearance of EGFP expression in treated cells. Sequence from PCR products corresponding to targeted site showed that insertion of a G into the exon of EGFP resulted in frame shift mutation. These results suggest that ZFN-mediated gene targeting can apply to caprine fetus fibroblasts, which may open a unique avenue toward the creation of gene knockout goats combining with somatic cell nuclear transfer.

Effectiveness of cultured Cordyceps sinensis combined with glucocorticosteroid on pulmonary fibrosis induced by bleomycin in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the treatment effects of cultured Cordyceps sinensis combined with glucocorticosteroid on experimental pulmonary fibrosis in rats induced by bleomycin.</p><p><b>METHOD</b>Fifty rats were randomly divided into five groups, including control group, model group, cultured C. sinensis groups, prednisone group, cultured C. sinensis combined with prednisone group. On experimental day 0, the rats were respectively intratracheally instilled with bleomycin, and rats in the control group and model group with the same volume of normal saline. One day after the injection, cultured C. sinensis and glucocorticosteroid was respectively given to rats daily by gastric gavage, while the same volume of normal saline was given to those in the control group and model group. On 28th d, bronchoalveolar lavage fluid (BALF) and lung tissue were collected. Histological changes of the lungs were evaluated by HE stain, Masson's trichrome stain. Collagen content of the lung tissue was assessed by hydroxyprolin concentration. Lung expression of CTGF protein was assessed by immunohistochemistry. The level of TGF-beta1 protein was measured by ELISA.</p><p><b>RESULT</b>Compared to model group, pulmonary fibrosis were alleviated in cultured C. sinensis and prednisone group, and CTGF expression, Hydroxyproline concentrations and protein TGF-beta1 were decreased. The combination effect of C. sinensis and prednisone group is augmented compared with using C. sinensis or prednisone group alone.</p><p><b>CONCLUSION</b>The cultured C. sinensis and prednisone alleviates pulmonary fibrosis, and the combination use of both drugs has synergia effects in anti-fibrous degeneration.</p>

Imaging findings of primary synodal sarcoma of the lung / 中华放射学杂志

r diagnosing the disease to combine pathology, immunohistochemistry and SYT-SSX gene detection.

Immunohistochemistry research of interface membrane around loosening hip prosthesis / 中国医师进修杂志

Objective Collecting the loosening periprosthetic interface-membrane, to discuss the mechanism of hip arthroplasty loosening. Methods The periprosthetic interface tissues of 29 hip arthroplasty revision cases from February 1995 to December 2003 were collected. The retrieved periprosthetic interface tissues were detected by immunohistochemistry. Some of them were studied by electronic microscope. Results (1)Transmission electronic microscope examination: the mitochondria swell. There were some substantia like lipid in the plasm of macrophages. Wear particles could be seen under scaning electronic microscope.(2)Immunohistochemistry: there were 22 IL-1? positive cases in cells of interface membrane. There were 29 IL-6 positive cases in cells of interface membrane. There were no positive results in TNF-? test. Conclusion (1)The wear particles of arthroplasty are important factors which cause biological reaction.(2)The interface membranes contain cytokine IL-1? and IL-6, which may play an important role in periprosthetic osteolysis and arthroplasty loosening.

Diagnosis and differential diagnosis of nodular lymphocyte-predominant Hodgkin's lymphoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the diagnosis and the differential diagnosis of nodular lymphocyte-predominant Hodgkin's lymphoma (NLPHL).</p><p><b>METHODS</b>245 cases of Hodgkin's lymphoma (HL) diagnosed between 1980 and 2000 from 3 hospitals in Guangzhou were reviewed. Four cases of NLPHL were confirmed according to the WHO classification of lymphoid neoplasms. Among the other 3 cases of NLPHL, 2 collected from other clinical centers and 1 from Fudan University Cancer Hospital. Immunohistochemistry (IHC) were performed on paraffin sections through SP technique using a panel of markers to define the large neoplastic cells (CD45, CD20, CD15, CD30 and vimentin) as well as the non-neoplastic background cells (CD3, CD20, CD45RO, CD57, CD68 and TIA-1).</p><p><b>RESULTS</b>Seven patients with NLPHL were 4 males and 3 females, age 29 to 70 years, average 43.8 years. All patients had lymphadenopathy. Histologically, in NLPHL, instead of the structure of normal lymph nodes, the tumor tissue became nodular in architecture. Characteristic lymphocytic and histiocytic (L&H) cells with scant cytoplasm and large multilobulated nuclei distributed among a predominant population of small lymphoid cells. The large cells exhibited a CD45+, CD20+, but CD15-, CD30- and vimentin-phenotype. The background cellularity was relatively rich in B cells and the majority of T-cells infiltrated were CD57(+) cells. TIA-1+ cells were few.</p><p><b>CONCLUSIONS</b>NLPHL can be diagnosed according to the morphologic and immunophenotypic features rather than by morphology alone. It is important to distinguish this tumor from its morphologic mimics, such as lymphocyte-rich classical Hodgkin's lymphoma (LRCHL) and T-cell rich B-cell lymphoma (TCRBCL). The immunophenotype of neoplastic cells and background cells are the helpful criteria for the differential diagnosis.</p>

Multilocular cystic renal clear cell carcinoma / 临床与实验病理学杂志

To study the clinical and pathological features and biologic behavior of multilocular cystic renal clear cell carcinoma(MLCRCCC). MethodsA case of MLCRCCC was stained with immunohistochemistry and the literature was reviewed. ResultsThe tumor was found in right kidney eighteen years before diagnosis. The majority of carcinoma cells showed cytokeratin,CEA and vimentin positivities by immunohistochemical staining. ConclusionMLCRCCC is a rare histologic subtype of renal cell carcinoma and usually cured by radical surgery. It originates from tubular epithelial cells of the kidney and appears to be a low - graded malignant neoplasm.