Objective:To investigate the clinicopathological factors affecting the prognosis of colorectal cancer patients with liver metastasis and how to select therapeutic method. Methods:The clinical records of 146 colorectal cancer patients with liver metastases who were admitted in Cancer Hospital of Guizhou Province from March 1997 to March 2007 were collected and made a retrospective analysis. The survival rate of the 146 patients was calculated by using life table method. Kaplan-Meier method and log-rank test were used for univariate analysis on clinicopathological features and therapeutic modalities. The multivariate analysis was performed by using COX regression model. The prognostic index (PI) of patients was calculated based on the result of multivariate analysis. The patients were classified into different risk groups according to PI value and the survival rate was compared between the different groups. Rusults:The overall 1-, 3-, and 5-year survival rates were 62.0%, 15.5%and 6.2%, respectively. Univariate analysis revealed that the following factors were related with the prognosis of colorectal cancer patients with liver metastasis. They included pathological classification, histological grade, serum CEA(carcinoembryonic antigen)level, primary tumor resection, local lymph node metastasis, number and size of liver metastases, distribution and initiation time of liver metastases, extrahepatic invasion, with or without surgery and che-motherapy for liver metastasis, and chemotherapeutic regimen selection. Multivariate analysis showed that the serum CEA level, extrahepatic metastasis, number and size of liver metastases, primary tumor resection, and chemotherapeutic regimen were independent prognostic factors for colorectal patients with liver metastasis.Conclusion:The therapeutic modality had an obvious effect on the prognosis of colorectal cancer with liver metastasis. Active treatment for primary tumor and metastatic lesions increased the survival rate of patients. Serum CEA levels, with or without extrahepatic metastases, and the number and size of liver metastases were prognostic factors. PI value could be used to predict the prognosis of colorectal cancer patients with liver metastasis.

OBJECTIVE To assess the prevalence and analyze the influence factors of laryngopharyngeal reflux disease(LPRD) in the Fuzhou region, in order to provide a theoretical basis for the development of prevention and control measures for LPRD. METHODS A questionnaire survey in residents in Fuzhou by a random cluster sampling was carried out. Individual information, reflux symptom index(RSI) of Belafsky and risk factors were included. Patients more than 13 scores of RSI were defined as LPRD. Data were statistically analyzed. RESULTS A total of 4100 residents were investigated, 4063 of them were available. The prevalence of LPRD was 5.00%. Often eating too much, often drinking strong tea, menolipsis, rhinitis, tonsillitis were closely related to LPRD. CONCLUSION The prevalence of LPRD in Fuzhou region were closely related to many factors.

Objective To investigate the feasibility of using recombinant infectious clones of hu-man coronavirus OC43 (HCoV-OC43) as a vector for the expression of exogenous genes and to analyze the insertion sites. Methods Based upon pBAC-OC43FL, a full-length cDNA infectious clone of HCoV-OC43, three recombinant expression plasmids ( pBAC-OC43-GFPΔNS2, pBAC-OC43-GFPΔNS12. 9 and pBAC-OC43-N-GFP) were respectively constructed by replacing NS2 and NS12. 9 genes with the reporter gene en-coding the green fluorescent protein ( GFP ) and inserting the reporter gene after the N gene by using the overlapping-PCR and in vitro ligation. Reverse genetics techniques were used for viral rescue. All of the res-cued virus strains were characterized by immunofluorescence assay ( IFA) and Western blot ( WB) assay af-ter transfecting BHK-21 cells with the recombinant viruses. Results Two recombinant viruses, OC43-GFPΔNS2 and OC43-GFPΔNS12. 9, could be successfully rescued by transfection the BHK-21 cells with pBAC-OC43-GFPΔNS2 and pBAC-OC43-GFPΔNS12. 9 plasmids. The expressed GFP was observed in BHK-21 cells transfected with pBAC-OC43-GFPΔNS2 or pBAC-OC43-GFPΔNS12. 9 plasmids, but not in the cells transfected with the pBAC-OC43-N-GFP plasmid. An efficient and stable expression of GFP was observed in the pBAC-OC43-GFPΔNS2 plasmid-transfected cells. The 10th generation of OC43-GFPΔNS2 virus was ob-tained after repeated freezing and thawing. The expression of GFP and N protein were detected in cells infec-ted with the OC43-GFPΔNS2 virus after 10 passages. Conclusion The NS2 gene of HCoV-OC43 could be used as a promising insertion site of the pBAC-OC43 FL infectious clone for the expression of exogenous genes. This study might provide a platform for further researches on the replication of HCoV-OC43 and the development of human coronavirus-based vectors.

To investigate the genetic character and origin of the first imported infection case of middle East respiratory syndrome coronavirus (named as MERS-CoV_China GD01), RNA was extracted from swabs of this patient followed by RT-PCR amplification. All coding gene of structural (S, E, M, E) and accessory (ORF3, ORF4a, ORF4b, ORF5, ORF8b) proteins were sequenced and analyzed. Phylogenetic analyses of structural protein coding genes of MERS-CoV_ China GD01 indicates that several substitutes exists in S coding gene and its origin belong group 5 of MERS-CoV, which were recent circulated in Saudi Arabia area, while other three structural genes (N, E, M) were very conserved. Phylogenetic analyses of accessory protein coding genes of MERS-CoV China GD01 indicates that several substitutes exists among ORF3, ORF4a, ORF4b and ORF5, while ORF8b was conserved. In conclusion, genome of MERS-CoV_ China GD01 was general conserved although several genetic variations were found among structural and accessory protein coding genes. This is the first report on sequencing and phylogenetic analyses of the first imported MERS case in China, which may pay the way for prevention and control of imported MERS-CoV infection.
China ; Conserved Sequence ; Coronavirus Infections ; transmission ; virology ; Evolution, Molecular ; Genomics ; Humans ; Middle East Respiratory Syndrome Coronavirus ; genetics ; physiology ; Phylogeny ; Sequence Analysis ; Viral Proteins ; genetics

OBJECTIVETo compare the modulatory effects of human amniotic epithelial cells (H-AECs), human amniotic mesenchymal cell (HA-MSCs), umbilical mesenchymal cells (UC-MSCs) on the inflammatory status of lipopolysaccharide (LPS)-induced RAW264.7 cells.

METHODSRAW264.7 cells stimulated with LPS were co-cultured with H-AECs, HA-MSCs, or UC-MSCs or cultured in conditioned media of the 3 stem cells to assess the changes of the inflammatory status of RAW264.7 cells. The migration ability, nitric oxide concentration, and expressions of the pro-inflammatory and anti-inflammatory genes, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNFα), and inducible nitric oxide synthase (NOS-2) of M1 macrophages, and Arg-1, CD206, and CD36 of M2 macrophages, were detected in the co-cultures.

RESULTSCompared with the control macrophages, RAW264.7 cells cultured in the conditioned media of H-AECs, HA-MSCs, and UC-MSCs all showed significantly lowered migration abilities (P<0.05). Co-culture with H-AECs, but not the other two stem cells, resulted in a significant reduction of NO production (P<0.05) and significant down-regulation of IL-1β, TNFα, NOS-2, and INFβ expressions in RAW264.7 cells; co-culture with HA-MSCs and UC-MSCs only caused a down-regulation of INFβ mRNA expression. In all the 3 RAW264.7 and stem cell co-cultures, the expressions of the inflammation related genes including Arg-1, CD206, and CD36 were up-regulated significantly.

CONCLUSIONH-AECs, HA-MSCs, and UC-MSCs can all prevent RAW264.7 cells from differentiating into M2 macrophages, but their effects and mechanisms are different from one another.

Adult Stem Cells ; cytology ; Animals ; Cell Line ; Coculture Techniques ; Culture Media, Conditioned ; Down-Regulation ; Humans ; Inflammation ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; Macrophages ; cytology ; Mesenchymal Stromal Cells ; cytology ; Mice ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective:To establish an HPLC method for the determination of paraquat in human serum.Methods:The analytical column was a Kromasil C18column (200mm×4.6mm,5μm).The mobile phase was a mixture of acetonitrile and water ( containing 0.03 mol·L-1sodium heptanesulfonate and 0.24 mol·L-1phosphoric acid) (3 :97,pH was adjusted to 2.0 by triethylamine),the detection wavelength was set at 258 nm,the column temperature was 25℃,the injection volume was 20 μl,and the flow rate was 0.8 ml·min-1.Results:The calibration curve of paraquat was linear within the range of 0.106-10.6 mg·L-1( r =0.999 3),and the lower limit of detection was 0.065 mg·L-1. The absolute recovery of paraquat at low,medium and high concentration was more than 89.4%,and the method recovery was more than 94.4%. The intra-day RSDs were 0.12%-1.74%,and the inter-day RSDs were 0.44%-2.89%.Conclusion:The method is simple,quick,accurate,sensitive and specific,and can be used for detecting paraquat con-centration in human serum.

Influenza poses a serious threat to global public health and causes serious economic los-ses. Vaccination is the most effective measure to prevent influenza. However, the mismatch between the vac-cine strain and the epidemic strain, which results from antigenic drift and antigen conversion of influenza vi-rus, often invalidates the conventional vaccine stockpiles. mRNA vaccine is a relatively safe platform com-posed of nucleic acid. Improvements in genetic engineering technology and novel delivery systems have great-ly promoted the research and development of mRNA vaccines against influenza. mRNA vaccines are promis-ing candidates for the rapid and efficient prevention of seasonal influenza epidemics and influenza pandemics.

Objective:To preliminarily observe the clinical efficacy of microwave hyperthermia combined with intensity-modulated radiotherapy (IMRT) and chemotherapy for patients with locally advanced gastric cancer.Methods:Forty patients who could not been operated or refused operation were enrolled in this clinical trial, who were confirmed as locally advanced proximal or distal gastric cancer by gastroscopy pathology and imaging. Radiotherapy was delivered by IMRT technology for 5 times per week with a total dose of 46 to 56 Gy (median dose of 50 Gy) in 25 to 28 fractions. Synchronous hyperthermia was given at 42 to 44℃ twice a week, 45 min/time. S-1 or capecitabine-based synchronous chemotherapy was performed, d1-14/3 weeks. The symptom remission rate, adverse reactions, objective remission rate (complete and partial remission) and survival were observed.Results:A total of 40 patients, aged between 56 and 83 years (median age of 71 years), were enrolled in this study. The male-to-female ratio was 7: 1. Among them, 38 cases (95%) showed symptom remission. The most common adverse reactions were grade 1-2 gastrointestinal reactions and leukopenia. The objective remission rate was 87.5%, the 2-year progression-free survival and overall survival rates were 68.6% and 70.5%, respectively.Conclusion:Preliminary findings demonstrate that microwave hyperthermia combined with chemoradiotherapy achieve satisfactory outcomes and yield tolerable toxicity in patients with locally advanced gastric cancer.

Objective:To establish and evaluate a rapid nucleic acid detection method for SARS-CoV-2 based on COYOTE ? Flash20 real-time fluorescent quantitative PCR instrument. Methods:A rapid reaction system was constructed by using specific primer and probe sets targeting ORF1ab and N gene of SARS-CoV-2, and the sensitivity and specificity of the system were verified. At the same time, 108 clinical samples of COVID-19 were used to evaluate the application of this method.Results:The detection method did not require nucleic acid extraction, and the manual operation time was only one minute. After the sample was sent to the system, the test could be completed in 30 minutes. The detection limit of this method was 4×10 2 copies/ml. It had no cross-reactivity with other human coronaviruses (including HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKU1, SARS-CoV and MERS-CoV) and other respiratory viruses. The evaluation of clinical sample application showed that the total coincidence rate with the conventional RT-qPCR which required nucleic acid extraction was 98.15%. Conclusions:Through the application evaluation of the rapid fluorescent quantitative PCR method of SARS-CoV-2, it was found that the method was simple, fast, specific and sensitive, and it was suitable for real-time and rapid detection needs in varieties of situations.

Objective To evaluate the expression profile of succinate dehydrogenase (SDH)B and SDHC in pheochromocytoma (PCC) and paraganglioma(PGL) (collectively abbreviated as PPGL), and their value in the early diagnosis of malignancy. Methods SDHB and SDHC immunohistochemistry were performed on 140 tumor specimens from 126 PPGL patients (PCC n＝62, PGL n＝61, PCC+PGL n＝3). Results (1) Germline mutation status of 67 patients were determined, of which, identifying 37(55.2%) patients with germline mutation: 2 (3.0%) SDHA, 18 ( 26. 9%) SDHB, 2 ( 3. 0%) SDHC, 5 ( 7. 5%) SDHD, 2 ( 3. 0%) VHL, 7 ( 10. 4%) RET, and 1(1.5%) NF1; and 30 (44.8%) individuals without known mutation. (2) Among 30 PPGLs from 27 patients with SDH-related (SDHx) mutations, 96.7%(29/30) stained negative for SDHB, 76.7%(23/30) stained negative for SDHC, while only 28.6%(14/49) and 18.4%(9/49) stained negative for SDHB and SDHC respectively in the 49 PPGLs without SDHx mutation (P<0.05). (3) The sensitivity of the SDH immunostaining in detecting the presence of germline SDHx mutation was 96.7%for SDHB and 76.7%for SDHC, while the specificity was 71.4%for SDHB and 81.6% for SDHC. ( 4 ) Among PPGLs without SDHB expression, 22. 9% were malignant. This percentage is significantly higher than that in PPGLs with preserved SDHB expression (3.8%, P<0.05). Conclusion SDHB and SDHC immunohistochemistry may serve as post-surgical screening tools to predict the presence of germline SDHx mutation in PPGLs. Negative SDHB expression calls for intense follow-up to rule out malignancy.