Objective To evaluate the inhibitory effect of Lactobacillus plantarum mouthwash on the progression of caries in isolated teeth.Methods Human isolated teeth were infected with Streptococcus mutans solution,and treated with saline,commercial mouth-wash,and Lactobacillus plantarum mouthwash,respectively,for 28 days.Tooth changes were recorded to assess their inhibitory effect on caries progression by light microscopy and scanning electron microscopy.Results Lactobacillus plantarum had an inhibitory effect on caries progression,with significant effects on deep dentinal caries in pit and fissure,and similar efficacy to commercial mouthwash on smooth surface caries.Calcium loss in dentin tubules was observed under an electron microscope,with Lactobacillus plantarum group less than commercial group and saline group.Conclusion Lactobacillus plantarum mouthwash can inhibit the growth of Streptococcus mutans and effectively prevent caries,particularly in controlling the progression of deep pit and fissure caries.

Objective To evaluate the inhibitory effect of Lactobacillus plantarum mouthwash on the progression of caries in isolated teeth.Methods Human isolated teeth were infected with Streptococcus mutans solution,and treated with saline,commercial mouth-wash,and Lactobacillus plantarum mouthwash,respectively,for 28 days.Tooth changes were recorded to assess their inhibitory effect on caries progression by light microscopy and scanning electron microscopy.Results Lactobacillus plantarum had an inhibitory effect on caries progression,with significant effects on deep dentinal caries in pit and fissure,and similar efficacy to commercial mouthwash on smooth surface caries.Calcium loss in dentin tubules was observed under an electron microscope,with Lactobacillus plantarum group less than commercial group and saline group.Conclusion Lactobacillus plantarum mouthwash can inhibit the growth of Streptococcus mutans and effectively prevent caries,particularly in controlling the progression of deep pit and fissure caries.

The local microenvironment is essential to stem cell-based therapy for ischemic stroke, and spatiotemporal changes of the microenvironment in the pathological process provide vital clues for understanding the therapeutic mechanisms. However, relevant studies on microenvironmental changes were mainly confined in the acute phase of stroke, and long-term changes remain unclear. This study aimed to investigate the microenvironmental changes in the subacute and chronic phases of ischemic stroke after stem cell transplantation. Herein, induced pluripotent stem cells (iPSCs) and neural stem cells (NSCs) were transplanted into the ischemic brain established by middle cerebral artery occlusion surgery. Positron emission tomography imaging and neurological tests were applied to evaluate the metabolic and neurofunctional alterations of rats transplanted with stem cells. Quantitative proteomics was employed to investigate the protein expression profiles in iPSCs-transplanted brain in the subacute and chronic phases of stroke. Compared with NSCs-transplanted rats, significantly increased glucose metabolism and neurofunctional scores were observed in iPSCs-transplanted rats. Subsequent proteomic data of iPSCs-transplanted rats identified a total of 39 differentially expressed proteins in the subacute and chronic phases, which are involved in various ischemic stroke-related biological processes, including neuronal survival, axonal remodeling, antioxidative stress, and mitochondrial function restoration. Taken together, our study indicated that iPSCs have a positive therapeutic effect in ischemic stroke and emphasized the wide-ranging microenvironmental changes in the subacute and chronic phases.
Animals ; Cell Differentiation ; Disease Models, Animal ; Ischemic Stroke ; Proteomics ; Rats ; Stem Cell Transplantation/methods* ; Stroke/therapy*

Objective: To investigate the principles of differential diagnosis of pulmonary infiltrates in cancer patients during the outbreak of novel coronavirus (2019-nCoV) by analyzing one case of lymphoma who presented pulmonary ground-glass opacities (GGO) after courses of chemotherapy. Methods: Baseline demographics and clinicopathological data of eligible patients were retrieved from medical records. Information of clinical manifestations, history of epidemiology, lab tests and chest CT scan images of visiting patients from February 13 to February 28 were collected. Literatures about pulmonary infiltrates in cancer patients were searched from databases including PUBMED, EMBASE and CNKI. Results: Among the 139 cancer patients underwent chest CT scans before chemotherapy, pulmonary infiltrates were identified in eight patients (5.8%), five of whom were characterized as GGOs in lungs. 2019-nCoV nuclear acid testing was performed in three patients and the results were negative. One case was a 66-year-old man diagnosed as non-Hodgkin lymphoma and underwent CHOP chemotherapy regimen. His chest CT scan image displayed multiple GGOs in lungs and the complete blood count showed decreased lymphocytes. This patient denied any contact with confirmed/suspected cases of 2019-nCoV infection and without fever and other respiratory symptoms. Considering the negative result of nuclear acid testing, this patient was presumptively diagnosed as viral pneumonia and an experiential anti-infection treatment had been prescribed for him. Conclusions The 2019 novel coronavirus disease (COVID-19) complicates the clinical scenario of pulmonary infiltrates in cancer patients. The epidemic history, clinical manifestation, CT scan image and lab test should be combined consideration. The 2019-nCoV nuclear acid testing might be applicated in more selected patients. Active anti-infection treatment and surveillance of patient condition should be initiated if infectious disease is considered.

Objective To study and search for a balance between the image quality and acquisition speed in tomography of whole body bone scan.Methods Adjustments of acquisition conditions were carried out gradualy every two months since April 2014. The qualities of fused SPECT/CT images were diagnosed by three doctors. Then the picture would be evaluated comprehensively by analyzing image quality and image resolution after adjusting image acquisition conditions.Results Seven kinds of image acquisition conditions taken were in line with diagnostic requirements. The third method is extended to clinical work best.Conclusion To obtain a high colection effi ciency, parameters of bone tomography acquisition can be set a frame of 5 seconds, total 64 (5.625o), automatic probe close and continuous scanning. Also recommends the use of "continuous" instead of "step and shoot" approach in bone SPECT acquisition. tomography, emission-computed, single-photon, bone tomography, program optimization.

OBJECTIVEThe aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens.

METHODSLung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical (IHC) ALK(D5F3) detecting ALK protein expression was performed in 203 prepared formalin-fixed paraffin-embedded (FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid (BL) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization (FISH). Six patients with ALK IHC-positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments: (1) Comparison of the results of 4% neutral buffered formalin fixed for different time (24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks; (2) Comparing qRT-PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens.

RESULTSAmong the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results (by at least one method), with an ALK test ratio of 90.4% (207/229). FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK (D5F3) were successfully performed in all the 203 FFPE cell blocks (100%), and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT-PCR, and 96 out of 98 (97.96%) cytologic samples were successfully performed.18 out of 19 IHC ALK-positive cases were verified to be of ALK fusion status by qRT-PCR. The concordance rate was 94.7% (Kappa=0.967, P<0.001) between Ventana IHC ALK (D5F3) and qRT-PCR, and the sensitivity of the Ventana IHC ALK (D5F3) assay compared with qRT-PCR was 100% and the specificity was 98.7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK (D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK (D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT-PCR test and ALK gene fusion showed good concordance.

CONCLUSIONSThe standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK (D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK (D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT-PCR may be an alternative option for the detection of ALK gene fusion.

Adenocarcinoma ; drug therapy ; enzymology ; genetics ; pathology ; Alkaline Phosphatase ; genetics ; metabolism ; Gene Fusion ; Genes, erbB-1 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; drug therapy ; enzymology ; genetics ; pathology ; Protein Kinase Inhibitors ; therapeutic use ; Proteomics ; Pyrazoles ; therapeutic use ; Pyridines ; therapeutic use ; Sensitivity and Specificity

Objective The aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens. Methods Lung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical ( IHC) ALK( D5F3) detecting ALK protein expression was performed in 203 prepared
formalin?fixed paraffin?embedded ( FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid ( BL ) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT?PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization ( FISH) . Six patients with ALK IHC?positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments:( 1) Comparison of the results of 4% neutral buffered formalin fixed for different time ( 24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks;(2) Comparing qRT?PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens. Results Among the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results ( by at least one method), with an ALK test ratio of 90.4% (207/229).FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK ( D5F3) were successfully performed in all the 203 FFPE cell blocks ( 100%) , and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT?PCR, and 96 out of 98 ( 97. 96%) cytologic samples were successfully performed.18 out of 19 IHC ALK?positive cases were verified to be of ALK fusion status by qRT?PCR. The concordance rate was 94.7% ( Kappa=0.967, P<0.001) between Ventana IHC ALK( D5F3) and qRT?PCR, and the sensitivity of the Ventana IHC ALK ( D5F3) assay compared with qRT?PCR was 100%and the specificity was 98. 7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK(D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK(D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT?PCR test and ALK gene fusion showed good concordance. Conclusions The standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK( D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK( D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT?PCR may be an alternative option for the detection of ALK gene fusion.

Objective The aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens. Methods Lung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical ( IHC) ALK( D5F3) detecting ALK protein expression was performed in 203 prepared
formalin?fixed paraffin?embedded ( FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid ( BL ) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT?PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization ( FISH) . Six patients with ALK IHC?positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments:( 1) Comparison of the results of 4% neutral buffered formalin fixed for different time ( 24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks;(2) Comparing qRT?PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens. Results Among the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results ( by at least one method), with an ALK test ratio of 90.4% (207/229).FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK ( D5F3) were successfully performed in all the 203 FFPE cell blocks ( 100%) , and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT?PCR, and 96 out of 98 ( 97. 96%) cytologic samples were successfully performed.18 out of 19 IHC ALK?positive cases were verified to be of ALK fusion status by qRT?PCR. The concordance rate was 94.7% ( Kappa=0.967, P<0.001) between Ventana IHC ALK( D5F3) and qRT?PCR, and the sensitivity of the Ventana IHC ALK ( D5F3) assay compared with qRT?PCR was 100%and the specificity was 98. 7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK(D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK(D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT?PCR test and ALK gene fusion showed good concordance. Conclusions The standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK( D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK( D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT?PCR may be an alternative option for the detection of ALK gene fusion.

OBJECTIVEThe aim of this study was to investigate the prevalence of Clostridium difficile (C. difficile) infection and the risk factors for acquisition of C. difficile-associated diarrhea (CDAD) among cancer patients who received chemotherapy or radiation therapy.

METHODSWe analyzed 277 stool samples from cancer patients with diarrhea between Sep 2010 and Dec 2011 in our hospital. Stool C. difficile toxin A/B test, stool culture for C. difficile and routine stool examination were performed. In addition, the risk factors for CDAD were investigated in a set of 41 C. difficile toxin-positive cancer patients and 82 matched C. difficile toxin-negative controls by univariate analysis and multivariate analysis.

RESULTSOut of a total of 277 cancer patients with diarrhea, 41 (14.8%) were C. difficile toxin-positive. Among these 41 cases, 11 (26.8%, 11/41) were C. difficile culture-positive. Univariate analysis showed that antibiotics use (P = 0.853), proton pump inhibitor use (P = 0.718), hypoproteinemia (P = 0.139) and white blood cell count (P = 0.454) did not appear to be associated with acquisition of CDAD in cancer patients. However, receiving chemotherapy (P = 0.023), receiving radiotherapy (P = 0.003), a positive fecal occult blood test result (P = 0.005) and the presence of fecal leukocytes (P = 0.007) showed close association with acquisition of CDAD in cancer patients. Multivariate analysis showed that receiving chemotherapy (OR, 8.308; 95% CI, 1.997-34.572; P = 0.004) and a positive result of fecal occult blood test (OR, 8.475; 95% CI, 1.463-49.109; P = 0.017) were independent risk factors for acquisition of CDAD among cancer patients.

CONCLUSIONSOur results support that receiving chemotherapy and a positive fecal occult blood test result are independent risk factors for acquisition of CDAD among cancer patients. Cancer patients who are at high-risk for CDAD should take stool C. difficile toxin A/B test and stool culture for C. difficile regularly and prevention of CDAD.

Clostridium difficile ; Diarrhea ; epidemiology ; microbiology ; Enterocolitis, Pseudomembranous ; epidemiology ; Humans ; Neoplasms ; epidemiology ; microbiology ; Risk Factors

OBJECTIVEThe aim of this study was to establish a standard protocol for detection of EGFR mutations in cytologic specimens.

METHODS287 cytologic samples were collected from the patients who were suspected of having lung cancer at six hospitals in Beijing. A detection protocol for EGFR mutations was designed. Two comparative experiments were carried out for the coincidence in EGFR mutation rates between direct sequencing (Seq) and amplification refractory mutation system (ARMS) methods, and between 40 matched cytologic samples with formaldehyde-fixed paraffin embedded (FFPE) cytologic blocks and cytospin slides.

RESULTSTumor cells were found in 236 out of 287 cases (82.2%, 236/287) . Among them, there were 31 cases (13.1%, 31/236) of low tumor cell content samples and 205 cases (86.9%, 205/236) of high tumor cell content samples. 180 cases in the high tumor cell content samples (87.8%, 180/205) were diagnosed to be consistent with NSCLC. 25 out of 194 cases were ruled out or indefinite to be diagnosed as NSCLC by immunohistochemistry. By direct sequencing, the mutation rate of EGFR was 27.8% (50/180) in NSCLC samples and 28.2% (50/177) in adenocarcinoma samples (high tumor content samples) . By ARMS, the mutation rate of EGFR was 45.6% (82/180) in NSCLC samples and 46.3% (82/177) in adenocarcinoma samples (high tumor content samples). The EGFR mutation rate in low tumor content samples was 38.7% (12/31) , there was no significant difference in EGFR mutation rates between the groups of low tumor cell content samples and high tumor cell content samples (P = 0.12). The concordance rate of EGFR mutation rates was 100% between scraping tumor cells from slides samples and from FFEP blocks in the 40 matched samples. Forty-eight out of 180 definitive NSCLC patients received Gefitinib therapy. The FPS was 12 months in the gefitinib-treated ARMS⁺ group and 2 months in the ARMS⁻ group (P < 0.001), and the OS was 19 months in the gefitinib-treated ARMS⁺ group and 7 months in the ARMS⁻ group (P = 0.003), but no significant differences were found in the efficacy (PFS and OS) of Gefitinib between Seq⁺ and Seq⁻ groups (P = 0.227, P = 0.510, respectively), and Seq⁺/ARMS⁺ and Seq⁻/ARMS⁺ groups (P = 0.354, P = 0.334, respectively).

CONCLUSIONSThe detection protocol for EGFR mutations in cytological specimens introduced in this study is tested to be reliable and feasible. Pathological evaluation and immunohistochemistry are important in the detection procedure of EGFR mutations in cytologic specimens. High sensitivity methods should be selected for detection of EGFR mutations in cytologic samples.

Adenocarcinoma ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Humans ; Lung Neoplasms ; diagnosis ; epidemiology ; metabolism ; Mutation ; Mutation Rate ; Polymerase Chain Reaction ; Receptor, Epidermal Growth Factor ; genetics ; metabolism