AIM: To evaluate the diagnostic potential of cytokeratin-19 (CK-19) mRNA for the detection of disseminated tumor cells in blood, bone marrow and peritoneal lavage in patients with ductal adenocarcinoma of the pancreas...AIM: To evaluate the diagnostic potential of cytokeratin-19 (CK-19) mRNA for the detection of disseminated tumor cells in blood, bone marrow and peritoneal lavage in patients with ductal adenocarcinoma of the pancreas. METHODS: Sixty-eight patients with pancreatic cancer (n = 37), chronic pancreatitis (n = 16), and non-pan- creatic benign surgical diseases (n = 15, control group) were included in the study. Venous blood was taken preoperatively, intraoperatively and at postoperative d 1 and 10. Preoperative bone marrow aspirates and peritoneal lavage taken before mobilization of the tumor were analyzed. All samples were evaluated for disseminated tumor cells by CK-19-specific nested-PCR and quantitative fluorogenic RT-PCR. RESULTS: CK-19 mRNA expression was increased in 24 (64%) blood samples and 11 (30%) of the peritoneal lavage samples in the patients with pancreatic cancer. In 15 (40%) of the patients with pancreatic cancer, disseminated tumor cells were detected in venous blood and bone marrow and/or peritoneal lavage. In the peritoneal lavage, the detection rates were correlated with the tumor size and the tumor differentiation. CK-19 levels were increased in pT3/T4 and moderately/poorly differentiated tumors (G2/G3). Pancreatic cancer patients with at least one CK-19 mRNA-positive sample showed a trend towards shorter survival. Pancreatic cancerpatients showed significantly increased detection rates of disseminated tumor cells in blood and peritoneal lavage compared to the controls and the patients with chronic pancreatitis. CONCLUSION: Disseminated tumor cells can be detected in patients with pancreatic ductal adenocar- cinoma by CK-19 fluorogenic RT-PCR. In peritoneal lavage, detection rate is correlated with tumor stage and differentiation. In the clinical use, CK-19 is suitable for the distinction between malignant and benign pancreatic disease in combination with other tumor-specific markers.展开更多
AIM:To evaluate the diagnostic role of serum RASSF1A promoter hypermethylation in gastric and colorectal ad- enocarcinoma. METHODS:Methylation-specific polymerase chain reac- tion (MSPCR) was used to examine the promo...AIM:To evaluate the diagnostic role of serum RASSF1A promoter hypermethylation in gastric and colorectal ad- enocarcinoma. METHODS:Methylation-specific polymerase chain reac- tion (MSPCR) was used to examine the promoter meth- ylation status of the serum RASSF1A gene in 47 gastric adenocarcinoma patients, 45 colorectal adenocarcinoma patients, 60 patients with benign gastrointestinal dis- ease (30 with benign gastric disease and 30 with benign colorectal disease), and 30 healthy donor controls. Apaired study of RASSF1A promoter methylation status in primary tumor, adjacent normal tissue, and postopera- tive serum were conducted in 25 gastric and colorectal adenocarcinoma patients who later were underwent sur- gical therapy. RESULTS:The frequencies of detection of serum RASSF1A promoter hypermethylation in gastric (34.0%) and colorectal (28.9%) adenocarcinoma patients were significantly higher than those in patients with benign gastric (3.3%) or colorectal (6.7%) disease or in healthy donors (0%) (P < 0.01). The methylation status of RASSF1A promoter in serum samples was consistent with that in paired primary tumors, and the MSPCR results for RASSF1A promoter methylation status in paired preoperative samples were consistent with those in postoperative serum samples. The serum RASSF1A promoter hypermethylation did not correlate with patient sex, age, tumor differentiation grade, surgical therapy, or serum carcinoembryonic antigen level. Although the serum RASSF1A promoter hypermethylation frequency tended to be higher in patients with distant metastases, there was no correlation between methylation status and metastasis. CONCLUSION:Aberrant CpG island methylation within the promoter region of RASSF1A is a promising biomark- er for gastric and colorectal cancer.展开更多
Objective Nucleostemin (NS) is a GTP-conjugated protein located in the nucleoli of stem cells and some cancer cells,and maintains cell self-renewal.We aimed to evaluate NS as a potential target for lung carcinoma gene...Objective Nucleostemin (NS) is a GTP-conjugated protein located in the nucleoli of stem cells and some cancer cells,and maintains cell self-renewal.We aimed to evaluate NS as a potential target for lung carcinoma gene therapy by investigating NS gene expression and its effect on A549 cell proliferation.Methods NS mRNA and protein expression in A549,HepG2,SMMC-7721,HeLa,and U251 cells was analyzed by RT-PCR and western blotting following transfection of NS siRNAs and negative control siRNA (NC).The effect on cell proliferation was also analyzed by MTT assays.Results NS mRNA and protein were both expressed in A549 cells and four other tumor cell lines;the relative expression levels were similar in all five cell lines.The three pairs of NS siRNA,either transfected alone or cotransfected into A549 cells,could effectively inhibit the expression of NS mRNA and protein.Moreover,the interference ratio showed an obvious concentration-dependent relationship.NS siRNA treatment resulted in significant inhibition of A549 cell proliferation by 35.7%.Conclusion NS gene was not only highly expressed but also played an important role in A549 cell proliferation.Thus,targeting of NS may be a promising novel strategy for the treatment of lung carcinoma.展开更多
Increasing evidence indicates that sialic acid plays an important role during nerve regeneration. Sialic acids can be modified in vitro as well as in vivo using metabolic oligosaccharide engineering of the N-acyl side...Increasing evidence indicates that sialic acid plays an important role during nerve regeneration. Sialic acids can be modified in vitro as well as in vivo using metabolic oligosaccharide engineering of the N-acyl side chain. N-Propionylmannosamine(Man NProp) increases neurite outgrowth and accelerates the reestablishment of functional synapses in vitro. We investigated the influence of systemic Man NProp application using a specific in vivo mouse model. Using mice expressing axonal fluorescent proteins, we quantified the extension of regenerating axons, the number of regenerating axons, the number of arborising axons and the number of branches per axon 5 days after injury. Sciatic nerves from non-expressing mice were grafted into those expressing yellow fluorescent protein. We began a twice-daily intraperitoneal application of either peracetylated Man NProp(200 mg/kg) or saline solution 5 days before injury, and continued it until nerve harvest(5 days after transection). Man NProp significantly increased the mean distance of axonal regeneration(2.49 mm vs. 1.53 mm; P < 0.005) and the number of arborizing axons(21% vs. 16%; P = 0.008) 5 days after sciatic nerve grafting. Man NProp did not affect the number of regenerating axons or the number of branches per arborizing axon. The biochemical glycoengineering of the N-acyl side chain of sialic acid might be a promising approach for improving peripheral nerve regeneration.展开更多
AIM: To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-α (PegIFN-α) and ribavirin treatment. METHODS: In a retrospective study w...AIM: To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-α (PegIFN-α) and ribavirin treatment. METHODS: In a retrospective study we evaluated a spectrum of predictors of relapse after PegIFN-α and ribavirin treatment in 86 CHC patients with ETR. Viral loads were determined with real-time reverse transcrip-tion polymerase chain reaction. Hepatitis C virus geno-typing was performed by sequencing analysis. Patients with genotype 1 were treated for 48 wk with 180 μg PegIFN-α2a or 1.5 μg/kg PegIFN-α2b once weekly plus ribavirin at a dosage of 1000 mg/d for those under 75 kg or 1200 mg/d for those over 75 kg. Patients with geno- types 2 and 3 were treated for 24 wk with 180 μgPegIFN-α2a or 1.5 μg/kg PegIFN-α2b once weekly plus ribavirin at a dosage of 800 mg/d. RESULTS: In all ETR patients, binary logistic regression analysis identif ied absence of complete early virological response (cEVR) (OR 27.07, 95% CI: 3.09-237.26, P < 0.005), serum alkaline phosphatase (ALP) levels prior to therapy < 75 U/L (OR: 6.16, 95% CI: 2.1-18.03, P < 0.001) and body mass index > 26 kg/m2 (OR: 8.27, 95% CI: 2.22-30.84, P < 0.005) as independent predictors of relapse. When cEVR patients were analyzed exclusively, ALP prior to therapy < 75 U/L remained the only predictor of relapse. CONCLUSION: Lower levels of ALP prior to, during and after therapy seem to be associated with a higher risk of relapse in CHC patients with ETR.展开更多
BACKGROUND Segmental intrahepatic cholestasis caused by transjugular intrahepatic portosystemic shunt(TIPS)(SIC-T),is a rare complication of this technique and only referred by case reports.Thus,we conducted a systema...BACKGROUND Segmental intrahepatic cholestasis caused by transjugular intrahepatic portosystemic shunt(TIPS)(SIC-T),is a rare complication of this technique and only referred by case reports.Thus,we conducted a systematic,retrospective analysis to provide evidence regarding prevalence and consequences of this TIPS-induced bile duct compression.AIM To assess prevalence and outcome of SIC-T in a large TIPS-cohort.METHODS In this retrospective cohort study,we screened the institutional databases for all consecutive patients that were treated by TIPS-placement or TIPS-revision between January 2005 and August 2013.We analyzed radiologic images for signs of biliary congestion.Cases that were indicative of SIC-T were reviewed by two independent radiologists and additional patient data was collected.Descriptive statistics of patient demographics,indications for TIPS and procedural details were registered.Logistic regression analysis was performed to identify predictors for the development of SIC-T.RESULTS We analyzed 135 cirrhotic patients who underwent TIPS(mean age 55 years,79%male gender).Etiology of cirrhosis was alcohol in most cases and indications for TIPS were mainly refractory ascites and recurrent variceal bleeding.TIPS revision was necessary in 31 patients.We identified 4 cases(2.9%)of SIC-T in direct proximity of the TIPS-stent.Diagnosis was confirmed by CT-scan,MRI or endoscopic retrograde cholangio pancreaticography(ERCP).In two patients TIPS was implanted via the right and in one through the medial hepatic vein.One patient received TIPS-prolongation by multiple revisions.Most patients were asymptomatic but one cholangitic abscess necessitated a transhepatic drain.Logistic regression analysis identified TIPS-placement other than from medial hepatic vein to right portal vein as risk factor(OR 21.0)for SIC-T.CONCLUSION SIC-T ads to(mostly late)complications in the interventional treatment of cirrhotic portal hypertensions and can lead to cholangitic abscesses.Patients,particularly with multiple interventions,should be screened for SIC-T.展开更多
Objective: To explore the possible biological function of human nuclear receptor hLRH-1 in tumorigenesis and progress of colon cancer. Methods: Plasmids pcDNA3-hLRH-1 were introduced into SW480 cells via lipofectamine...Objective: To explore the possible biological function of human nuclear receptor hLRH-1 in tumorigenesis and progress of colon cancer. Methods: Plasmids pcDNA3-hLRH-1 were introduced into SW480 cells via lipofectamine. The expression of mRNA and protein of exogenous hLRH-1 were detected by RT-PCR and western blotting, respectively. MTT assay was carried out to survey the proliferation of SW480 cells with overexpression of hLRH-1. Meanwhile, the expression of proliferation-related genes cyclin E1 and cyclin D1, and apoptosis-related genes PTEN and Rb1, were analyzed by real- time RT-PCR. Results: The proliferation of SW480 cells was promoted under the condition of overexpression of hLRH-1. The expression of cyclin E1 was up-regulated significantly, while that of PTEN and Rb1 were down-regulated in SW480 cells with overexpressed hLRH-1. Conclusion: The expression of exogenous hLRH-1 in SW480 cells induced the proliferation resulting form up-regulation of cyclin E1, as well as participated in the regulation of apoptosis via influencing the expression of PTEN and Rb1.展开更多
Retinitis pigmentosa(RP)is an inherited retinal degenerative disease that begins with defective rod photoreceptor function,followed by impaired cone function,and complete blindness in its late stage.To date,however,th...Retinitis pigmentosa(RP)is an inherited retinal degenerative disease that begins with defective rod photoreceptor function,followed by impaired cone function,and complete blindness in its late stage.To date,however,there is no effective treatment for RP.By carrying a nonsense mutation in the Pde6b gene,rd1 mice display elevated cGMP in conjunction with higher intracellular Ca2+in their rod photoreceptors,resulting in fast retinal degeneration.Ca2+has been linked to activation of the mammalian target of rapamycin(mTOR)pathway.The mTOR pathway integrates extracellular and intracellular signals to sense the supply of nutrients and plays a central role in regulating protein and lipid synthesis as well as apoptosis and autophagy.In the present study,we showed that mTOR and phosphorylated mTOR(p-mTOR,activated form of mTOR)are up-regulated in rd1 photoreceptors at postnatal day 10(P10),a pre-degenerative stage.Moreover,the downstream effectors of mTOR,such as pS6K and S6K,are also increased,suggesting activation of the mTOR signaling pathway.Intravitreal administration of rapamycin,a negative regulator of mTOR,inhibits the mTOR pathway in rd1 photoreceptors.Consequently,the progression of retinal degeneration is slower and retinal function is enhanced,possibly mediated by activation of autophagy in the photoreceptors.Taken together,these results highlight rapamycin as a potential therapeutic avenue for retinal degeneration.展开更多
AIM:To investigate the effects of dexamethasone(DEX) and 1-(5-isoquinolinesulfonyl)-homopiperazine(HA1077) on actin cytoskeleton and β-catenin in cultured human trabecular meshwork(HTM) cells.METHODS: The HTM cells w...AIM:To investigate the effects of dexamethasone(DEX) and 1-(5-isoquinolinesulfonyl)-homopiperazine(HA1077) on actin cytoskeleton and β-catenin in cultured human trabecular meshwork(HTM) cells.METHODS: The HTM cells were separated from human eyeball and cultured in vitro.They were divided into control group,DEX(1×10^(-6)mol/L) group,HA1077(3×10^(-5)mol/L)group,and DEX(1×10^(-6)mol/L) and HA1077(3×10^(-5)mol/L)group.Actin cytoskeleton and β-catenin in HTM cells of the four groups were examined by immunofluorescence and Western blot analyses.RESULTS: In DEX group,there were reorganization of actin cytoskeleton and formation of cross linked actin networks(CLANs),which were partially reversed in DEX and HA1077 group.DEX treatment also induced an increased expression of β-catenin,which was obviously reduced in DEX and HA1077 group.Meanwhile,the cultured HTM cells in HA1077 group had lower expression of β-catenin than that in the control group. CONCLUSION: Our results show that HA1077 can reverse the changes of actin organization and expression of β-catenin induced by DEX in cultured HTM cells,suggesting that HA1077 may play an important role in increasing outflow and reducing intraocular pressure.展开更多
An outbreak of a novel coronavirus was reported in Wuhan,China,in late 2019.It has spread rapidly through China and many other countries,causing a global pandemic.Since February 2020,over 28 countries/regions have rep...An outbreak of a novel coronavirus was reported in Wuhan,China,in late 2019.It has spread rapidly through China and many other countries,causing a global pandemic.Since February 2020,over 28 countries/regions have reported confirmed cases.Individuals with the infection known as coronavirus disease-19(COVID-19)have similar clinical features as severe acute respiratory syndrome first encountered 17 years ago,with fever,cough,and upper airway congestion,along with high production of proinflammatory cytokines(PICs),which form a cytokine storm.PICs induced by COVID-19 include interleukin(IL)-6,IL-17,and monocyte chemoattractant protein-1.The production of cytokines is regulated by activated nuclear factor-kB and involves downstream pathways such as Janus kinase/signal transducers and activators transcription.Protein expression is also regulated by post-translational modification of chromosomal markers.Lysine residues in the peptide tails stretching out from the core of histones bind the sequence upstream of the coding portion of genomic DNA.Covalent modification,particularly methylation,activates or represses gene transcription.PICs have been reported to be induced by histone modification and stimulate exudation of hyaluronic acid,which is implicated in the occurrence of COVID-19.These findings indicate the impact of the expression of PICs on the pathogenesis and therapeutic targeting of COVID-19.展开更多
Background: Gastrointestinal symptoms are not rare among coronavirus disease 2019(COVID-19) patients, but there have been no reports regarding convalescent plasma therapy for the recovery of gastrointestinal problems ...Background: Gastrointestinal symptoms are not rare among coronavirus disease 2019(COVID-19) patients, but there have been no reports regarding convalescent plasma therapy for the recovery of gastrointestinal problems in COVID-19 patients.Case presentation: We present two cases of patients with COVID-19-associated recurrent diarrhea and positive fecal occult blood who successfully recovered after a one-time convalescent plasma administration.Conclusion: When COVID-19 patients develop recurrent or refractory gastrointestinal symptoms and fail to respond to the available treatment, alternative therapy with convalescent plasma administration may be considered.展开更多
Background: Telmisartan is a highly variable drug which is used to treat hypertension. This study compared to compare the bioavailability of two 40 mg Telmisartan tablets in adult and healthy Bangladeshi subjects. Mat...Background: Telmisartan is a highly variable drug which is used to treat hypertension. This study compared to compare the bioavailability of two 40 mg Telmisartan tablets in adult and healthy Bangladeshi subjects. Materials and Method: This study was open label, randomized, laboratory blind, single dose, three periods, two treatments, three-sequence, partial-replicate, crossover and comparative oral bioavailability study. In this study, 18 Bangladeshi subjects were enrolled and 17 subjects were completed. Serial blood samples were collected up to 96 hours following drug administration. By using Liquid Chromatography Mass Spectrometry (LC-MS/MS) method, plasma concentrations of Telmisartan were determined. Results: The two formulations of Telmisartan were considered bioequivalent if 90% Confidence Interval (CI) fall within the range of 80.00% - 125.00% for AUC parameters and reference-scaled-average bioequivalence of 69.84% - 143.19% for C<sub>max</sub>. The 90% Confidence Interval for C<sub>max</sub>, AUC<sub>0-t</sub> & AUC<sub>0-∞</sub> was found 84.88% - 107.79%, 89.76% - 109.55%, and 91.20% - 114.52%, respectively. Conclusion: According to rate and extent of absorption with the single dose of Reference Product (R): Micardis<sup><sup><sup>®</sup></sup></sup> 40 mg Tablet, under fasting conditions, a single dose of Telmisartan 40 mg Tablet is bioequivalent.展开更多
We present a new method of treatment for nasal valve dysfunction caused by insufficiency or stenosis in a patient who refused open septum revision despite a significant degree of septum deviation. The Anterior Spreade...We present a new method of treatment for nasal valve dysfunction caused by insufficiency or stenosis in a patient who refused open septum revision despite a significant degree of septum deviation. The Anterior Spreader Flap (ASF) technique was suggested as an alternative to open nasal septum revision and was performed under local anesthesia. Computational fluid dynamics (CFD) tests were performed pre- and post-operatively and our patient was asked to complete a self-assessment using a Visual Analog Scale (VAS) for nasal breathing (0 = free nasal breathing, 10 = complete nasal blockage) before and 12 months after surgery. The ASF is a minimally invasive endonasal procedure in which the caudal edge of the upper lateral cartilage is dissected from the septum, and folded inwards and fixed. The ASF allows for less airflow resistance and more free space. CFD techniques revealed a reduction in local pressure based on extended space. In addition, VAS scores improved from 9 to 2 points (right side) and from 8 to 2 points (left side). On the basis of these findings, the ASF technique can be considered a safe, minimally invasive spreader flap technique. It can easily be combined with other nasal surgical techniques, as it is necessary in most cases. In selected cases, the ASF may be performed as a single procedure in patients with nasal valve dysfunction caused by septum deviation as an alternative to open septum revision.展开更多
Objective To investigate the clinical application of Real-Time PCR for rapid detection of methicillin-resistant Staphylococcus aureus(MRSA) directly from nasopharyngeal swab specimens.Methods We collected the nasal an...Objective To investigate the clinical application of Real-Time PCR for rapid detection of methicillin-resistant Staphylococcus aureus(MRSA) directly from nasopharyngeal swab specimens.Methods We collected the nasal and throat swab specimens from patients or medical staffs in 3 intensive care units,blood laminar flow ward and respiratory ward in Beijing Hospital,Ministry of Health from December 2010to April 2011.Each sample was tested by RT-PCR and conventional culture-based method for the presence of MRSA.Results The total number of the specimens was 206.Compared with the conventional culture-based method,we demonstrated the diagnostic values for Real-Time PCR were 96.4%sensitivity,96.6%specificity,81.8%positive predictive rate,and 99.4%negative predictive rate.And the limit of detection was 10~2CFU/ml.Conclusions This Real-Time PCR is a simple,rapid,sensitive and specific method.With the high negative predictive value,it can be used for the exclusion of MRSA colonization or infection.However,the application of its low positive predictive value should be further evaluated.展开更多
Novel coronavirus,SARS-CoV-2 is responsible for causing a pandemic that has affected individuals worldwide,over 192 million people and about 4.1 million people died so far.The spread is ongoing and the numbers are sti...Novel coronavirus,SARS-CoV-2 is responsible for causing a pandemic that has affected individuals worldwide,over 192 million people and about 4.1 million people died so far.The spread is ongoing and the numbers are still increasing.Numerous therapeutic approaches have been explored and developed during this pandemic.Immunotherapy with virus-specific antibodies in convalescent plasma(CP)has shown potential benefits for various pathogenic diseases.In many instances,it is the only available and safe management option for the COVID-19 patients.Here we describe two confirmed cases of COVID-19 from two different geographical areas that were managed with standard treatment modalities initially.Both of the patients were presented with high-grade fever,dry cough,and sore throat.Lab reports showed increased values of D-dimer,serum ferritin,leukocyte count(LC),Lactate dehydrogenase(LDH),and C-reactive protein(CRP).Chest X-ray showed bilateral infiltration(multifocal and bilateral ground-glass opacities and consolidations with peripheral and basal predominance),consistent with the previous reports on COVID-19 infection.The patients received conservative treatment according to the hospital's protocol.The convalescent plasma(from recovered patients)infusion was the last treatment given to both patients.After the convalescent plasma transfusion,both patients showed a reduction of viral load,an increase of anti-SARS-CoV-2 IgG and IgM antibodies,reduction in lung infiltration,with no adverse events.However,further randomized controlled trials are needed to investigate the full scope of safety and efficacy(both short and long-term)of convalescent plasma therapy for COVID-19 and other related infections.展开更多
Ecological studies suggested a link between air pollution and severe COVID-19 outcomes,while studies accounting for individual-level characteristics are limited.In the present study,we aimed to investigate the impact ...Ecological studies suggested a link between air pollution and severe COVID-19 outcomes,while studies accounting for individual-level characteristics are limited.In the present study,we aimed to investigate the impact of short-term ambient air pollution exposure on disease severity among a cohort of 569 laboratory confirmed COVID-19 patients admitted to designated hospitals in Zhejiang province,China,from January 17 to March 3,2020,and elucidate the possible biological processes involved using transcriptomics.Compared with mild cases,severe cases had higher proportion of medical conditions as well as unfavorable results in most of the laboratory tests,and manifested higher air pollution exposure levels.Higher exposure to air pollutants was associated with increased risk of severe COVID-19 with odds ratio(OR)of 1.89(95%confidence interval(CI):1.01,3.53),2.35(95%CI:1.20,4.61),2.87(95%CI:1.68,4.91),and 2.01(95%CI:1.10,3.69)for PM_(2.5),PM_(10),NO_(2)and CO,respectively.OR for NO_(2)remained significant in two-pollutant models after adjusting for other pollutants.Transcriptional analysis showed 884 differentially expressed genes which mainly were enriched in virus clearance related biological processes between patients with high and low NO_(2)exposure levels,indicating that compromised immune response might be a potential underlying mechanistic pathway.These findings highlight the impact of shortterm air pollution exposure,particularly for NO_(2),on COVID-19 severity,and emphasize the significance in mitigating the COVID-19 burden of commitments to improve air quality.展开更多
文摘AIM: To evaluate the diagnostic potential of cytokeratin-19 (CK-19) mRNA for the detection of disseminated tumor cells in blood, bone marrow and peritoneal lavage in patients with ductal adenocarcinoma of the pancreas. METHODS: Sixty-eight patients with pancreatic cancer (n = 37), chronic pancreatitis (n = 16), and non-pan- creatic benign surgical diseases (n = 15, control group) were included in the study. Venous blood was taken preoperatively, intraoperatively and at postoperative d 1 and 10. Preoperative bone marrow aspirates and peritoneal lavage taken before mobilization of the tumor were analyzed. All samples were evaluated for disseminated tumor cells by CK-19-specific nested-PCR and quantitative fluorogenic RT-PCR. RESULTS: CK-19 mRNA expression was increased in 24 (64%) blood samples and 11 (30%) of the peritoneal lavage samples in the patients with pancreatic cancer. In 15 (40%) of the patients with pancreatic cancer, disseminated tumor cells were detected in venous blood and bone marrow and/or peritoneal lavage. In the peritoneal lavage, the detection rates were correlated with the tumor size and the tumor differentiation. CK-19 levels were increased in pT3/T4 and moderately/poorly differentiated tumors (G2/G3). Pancreatic cancer patients with at least one CK-19 mRNA-positive sample showed a trend towards shorter survival. Pancreatic cancerpatients showed significantly increased detection rates of disseminated tumor cells in blood and peritoneal lavage compared to the controls and the patients with chronic pancreatitis. CONCLUSION: Disseminated tumor cells can be detected in patients with pancreatic ductal adenocar- cinoma by CK-19 fluorogenic RT-PCR. In peritoneal lavage, detection rate is correlated with tumor stage and differentiation. In the clinical use, CK-19 is suitable for the distinction between malignant and benign pancreatic disease in combination with other tumor-specific markers.
文摘AIM:To evaluate the diagnostic role of serum RASSF1A promoter hypermethylation in gastric and colorectal ad- enocarcinoma. METHODS:Methylation-specific polymerase chain reac- tion (MSPCR) was used to examine the promoter meth- ylation status of the serum RASSF1A gene in 47 gastric adenocarcinoma patients, 45 colorectal adenocarcinoma patients, 60 patients with benign gastrointestinal dis- ease (30 with benign gastric disease and 30 with benign colorectal disease), and 30 healthy donor controls. Apaired study of RASSF1A promoter methylation status in primary tumor, adjacent normal tissue, and postopera- tive serum were conducted in 25 gastric and colorectal adenocarcinoma patients who later were underwent sur- gical therapy. RESULTS:The frequencies of detection of serum RASSF1A promoter hypermethylation in gastric (34.0%) and colorectal (28.9%) adenocarcinoma patients were significantly higher than those in patients with benign gastric (3.3%) or colorectal (6.7%) disease or in healthy donors (0%) (P < 0.01). The methylation status of RASSF1A promoter in serum samples was consistent with that in paired primary tumors, and the MSPCR results for RASSF1A promoter methylation status in paired preoperative samples were consistent with those in postoperative serum samples. The serum RASSF1A promoter hypermethylation did not correlate with patient sex, age, tumor differentiation grade, surgical therapy, or serum carcinoembryonic antigen level. Although the serum RASSF1A promoter hypermethylation frequency tended to be higher in patients with distant metastases, there was no correlation between methylation status and metastasis. CONCLUSION:Aberrant CpG island methylation within the promoter region of RASSF1A is a promising biomark- er for gastric and colorectal cancer.
基金supported by the Plan of Promoting Sichuan University science research start up fund (NO.0082204127092)
文摘Objective Nucleostemin (NS) is a GTP-conjugated protein located in the nucleoli of stem cells and some cancer cells,and maintains cell self-renewal.We aimed to evaluate NS as a potential target for lung carcinoma gene therapy by investigating NS gene expression and its effect on A549 cell proliferation.Methods NS mRNA and protein expression in A549,HepG2,SMMC-7721,HeLa,and U251 cells was analyzed by RT-PCR and western blotting following transfection of NS siRNAs and negative control siRNA (NC).The effect on cell proliferation was also analyzed by MTT assays.Results NS mRNA and protein were both expressed in A549 cells and four other tumor cell lines;the relative expression levels were similar in all five cell lines.The three pairs of NS siRNA,either transfected alone or cotransfected into A549 cells,could effectively inhibit the expression of NS mRNA and protein.Moreover,the interference ratio showed an obvious concentration-dependent relationship.NS siRNA treatment resulted in significant inhibition of A549 cell proliferation by 35.7%.Conclusion NS gene was not only highly expressed but also played an important role in A549 cell proliferation.Thus,targeting of NS may be a promising novel strategy for the treatment of lung carcinoma.
文摘Increasing evidence indicates that sialic acid plays an important role during nerve regeneration. Sialic acids can be modified in vitro as well as in vivo using metabolic oligosaccharide engineering of the N-acyl side chain. N-Propionylmannosamine(Man NProp) increases neurite outgrowth and accelerates the reestablishment of functional synapses in vitro. We investigated the influence of systemic Man NProp application using a specific in vivo mouse model. Using mice expressing axonal fluorescent proteins, we quantified the extension of regenerating axons, the number of regenerating axons, the number of arborising axons and the number of branches per axon 5 days after injury. Sciatic nerves from non-expressing mice were grafted into those expressing yellow fluorescent protein. We began a twice-daily intraperitoneal application of either peracetylated Man NProp(200 mg/kg) or saline solution 5 days before injury, and continued it until nerve harvest(5 days after transection). Man NProp significantly increased the mean distance of axonal regeneration(2.49 mm vs. 1.53 mm; P < 0.005) and the number of arborizing axons(21% vs. 16%; P = 0.008) 5 days after sciatic nerve grafting. Man NProp did not affect the number of regenerating axons or the number of branches per arborizing axon. The biochemical glycoengineering of the N-acyl side chain of sialic acid might be a promising approach for improving peripheral nerve regeneration.
文摘AIM: To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-α (PegIFN-α) and ribavirin treatment. METHODS: In a retrospective study we evaluated a spectrum of predictors of relapse after PegIFN-α and ribavirin treatment in 86 CHC patients with ETR. Viral loads were determined with real-time reverse transcrip-tion polymerase chain reaction. Hepatitis C virus geno-typing was performed by sequencing analysis. Patients with genotype 1 were treated for 48 wk with 180 μg PegIFN-α2a or 1.5 μg/kg PegIFN-α2b once weekly plus ribavirin at a dosage of 1000 mg/d for those under 75 kg or 1200 mg/d for those over 75 kg. Patients with geno- types 2 and 3 were treated for 24 wk with 180 μgPegIFN-α2a or 1.5 μg/kg PegIFN-α2b once weekly plus ribavirin at a dosage of 800 mg/d. RESULTS: In all ETR patients, binary logistic regression analysis identif ied absence of complete early virological response (cEVR) (OR 27.07, 95% CI: 3.09-237.26, P < 0.005), serum alkaline phosphatase (ALP) levels prior to therapy < 75 U/L (OR: 6.16, 95% CI: 2.1-18.03, P < 0.001) and body mass index > 26 kg/m2 (OR: 8.27, 95% CI: 2.22-30.84, P < 0.005) as independent predictors of relapse. When cEVR patients were analyzed exclusively, ALP prior to therapy < 75 U/L remained the only predictor of relapse. CONCLUSION: Lower levels of ALP prior to, during and after therapy seem to be associated with a higher risk of relapse in CHC patients with ETR.
文摘BACKGROUND Segmental intrahepatic cholestasis caused by transjugular intrahepatic portosystemic shunt(TIPS)(SIC-T),is a rare complication of this technique and only referred by case reports.Thus,we conducted a systematic,retrospective analysis to provide evidence regarding prevalence and consequences of this TIPS-induced bile duct compression.AIM To assess prevalence and outcome of SIC-T in a large TIPS-cohort.METHODS In this retrospective cohort study,we screened the institutional databases for all consecutive patients that were treated by TIPS-placement or TIPS-revision between January 2005 and August 2013.We analyzed radiologic images for signs of biliary congestion.Cases that were indicative of SIC-T were reviewed by two independent radiologists and additional patient data was collected.Descriptive statistics of patient demographics,indications for TIPS and procedural details were registered.Logistic regression analysis was performed to identify predictors for the development of SIC-T.RESULTS We analyzed 135 cirrhotic patients who underwent TIPS(mean age 55 years,79%male gender).Etiology of cirrhosis was alcohol in most cases and indications for TIPS were mainly refractory ascites and recurrent variceal bleeding.TIPS revision was necessary in 31 patients.We identified 4 cases(2.9%)of SIC-T in direct proximity of the TIPS-stent.Diagnosis was confirmed by CT-scan,MRI or endoscopic retrograde cholangio pancreaticography(ERCP).In two patients TIPS was implanted via the right and in one through the medial hepatic vein.One patient received TIPS-prolongation by multiple revisions.Most patients were asymptomatic but one cholangitic abscess necessitated a transhepatic drain.Logistic regression analysis identified TIPS-placement other than from medial hepatic vein to right portal vein as risk factor(OR 21.0)for SIC-T.CONCLUSION SIC-T ads to(mostly late)complications in the interventional treatment of cirrhotic portal hypertensions and can lead to cholangitic abscesses.Patients,particularly with multiple interventions,should be screened for SIC-T.
基金the Young Scientific and Technical Innovation Foundation of Fujian Province (No. 2004J067)Foundation of Fuzhou General Hospital (No. 200638)
文摘Objective: To explore the possible biological function of human nuclear receptor hLRH-1 in tumorigenesis and progress of colon cancer. Methods: Plasmids pcDNA3-hLRH-1 were introduced into SW480 cells via lipofectamine. The expression of mRNA and protein of exogenous hLRH-1 were detected by RT-PCR and western blotting, respectively. MTT assay was carried out to survey the proliferation of SW480 cells with overexpression of hLRH-1. Meanwhile, the expression of proliferation-related genes cyclin E1 and cyclin D1, and apoptosis-related genes PTEN and Rb1, were analyzed by real- time RT-PCR. Results: The proliferation of SW480 cells was promoted under the condition of overexpression of hLRH-1. The expression of cyclin E1 was up-regulated significantly, while that of PTEN and Rb1 were down-regulated in SW480 cells with overexpressed hLRH-1. Conclusion: The expression of exogenous hLRH-1 in SW480 cells induced the proliferation resulting form up-regulation of cyclin E1, as well as participated in the regulation of apoptosis via influencing the expression of PTEN and Rb1.
基金supported by the National Precision Medicine Project(2016YFC0905200)(Z.Y.&H.Z.)National Natural Science Foundation of China(81570882(H.Z.),81770935(H.Z.))+1 种基金Department of Science and Technology of Sichuan Province,China((2020YJ0445)(H.Z.),2020ZYD037(Z.Y))CAMS Innovation Fund for Medical Sciences(2019-I2M-5-032)(Z.Y.)。
文摘Retinitis pigmentosa(RP)is an inherited retinal degenerative disease that begins with defective rod photoreceptor function,followed by impaired cone function,and complete blindness in its late stage.To date,however,there is no effective treatment for RP.By carrying a nonsense mutation in the Pde6b gene,rd1 mice display elevated cGMP in conjunction with higher intracellular Ca2+in their rod photoreceptors,resulting in fast retinal degeneration.Ca2+has been linked to activation of the mammalian target of rapamycin(mTOR)pathway.The mTOR pathway integrates extracellular and intracellular signals to sense the supply of nutrients and plays a central role in regulating protein and lipid synthesis as well as apoptosis and autophagy.In the present study,we showed that mTOR and phosphorylated mTOR(p-mTOR,activated form of mTOR)are up-regulated in rd1 photoreceptors at postnatal day 10(P10),a pre-degenerative stage.Moreover,the downstream effectors of mTOR,such as pS6K and S6K,are also increased,suggesting activation of the mTOR signaling pathway.Intravitreal administration of rapamycin,a negative regulator of mTOR,inhibits the mTOR pathway in rd1 photoreceptors.Consequently,the progression of retinal degeneration is slower and retinal function is enhanced,possibly mediated by activation of autophagy in the photoreceptors.Taken together,these results highlight rapamycin as a potential therapeutic avenue for retinal degeneration.
基金Supported by the Natural Science Foundation of China (No.81300768 No.81371048+4 种基金 No.81670853)Science and Technology Department of Sichuan Province of China (No.2015HH0031)Health and Family Planning Commission of Sichuan Province of China (No.100539 No. 090505 No.090514)
文摘AIM:To investigate the effects of dexamethasone(DEX) and 1-(5-isoquinolinesulfonyl)-homopiperazine(HA1077) on actin cytoskeleton and β-catenin in cultured human trabecular meshwork(HTM) cells.METHODS: The HTM cells were separated from human eyeball and cultured in vitro.They were divided into control group,DEX(1×10^(-6)mol/L) group,HA1077(3×10^(-5)mol/L)group,and DEX(1×10^(-6)mol/L) and HA1077(3×10^(-5)mol/L)group.Actin cytoskeleton and β-catenin in HTM cells of the four groups were examined by immunofluorescence and Western blot analyses.RESULTS: In DEX group,there were reorganization of actin cytoskeleton and formation of cross linked actin networks(CLANs),which were partially reversed in DEX and HA1077 group.DEX treatment also induced an increased expression of β-catenin,which was obviously reduced in DEX and HA1077 group.Meanwhile,the cultured HTM cells in HA1077 group had lower expression of β-catenin than that in the control group. CONCLUSION: Our results show that HA1077 can reverse the changes of actin organization and expression of β-catenin induced by DEX in cultured HTM cells,suggesting that HA1077 may play an important role in increasing outflow and reducing intraocular pressure.
基金Medical Scientific Research Foundation of Guangdong Province of China,Guangdong Provincial Health Commission,China,No.A2018356。
文摘An outbreak of a novel coronavirus was reported in Wuhan,China,in late 2019.It has spread rapidly through China and many other countries,causing a global pandemic.Since February 2020,over 28 countries/regions have reported confirmed cases.Individuals with the infection known as coronavirus disease-19(COVID-19)have similar clinical features as severe acute respiratory syndrome first encountered 17 years ago,with fever,cough,and upper airway congestion,along with high production of proinflammatory cytokines(PICs),which form a cytokine storm.PICs induced by COVID-19 include interleukin(IL)-6,IL-17,and monocyte chemoattractant protein-1.The production of cytokines is regulated by activated nuclear factor-kB and involves downstream pathways such as Janus kinase/signal transducers and activators transcription.Protein expression is also regulated by post-translational modification of chromosomal markers.Lysine residues in the peptide tails stretching out from the core of histones bind the sequence upstream of the coding portion of genomic DNA.Covalent modification,particularly methylation,activates or represses gene transcription.PICs have been reported to be induced by histone modification and stimulate exudation of hyaluronic acid,which is implicated in the occurrence of COVID-19.These findings indicate the impact of the expression of PICs on the pathogenesis and therapeutic targeting of COVID-19.
基金supported by the Key Foundation of Wuhan Huoshenshan Hospital (2020 [18])Key Research&Development Program of Jiangsu Province (BE2018713)+3 种基金Medical Innovation Project of Logistics Service (18JS005)the Foundation of Jiangsu Population Association (JSPA2019017)Medical Science and Technology Development FoundationNanjing Department of Health (YKK18179)。
文摘Background: Gastrointestinal symptoms are not rare among coronavirus disease 2019(COVID-19) patients, but there have been no reports regarding convalescent plasma therapy for the recovery of gastrointestinal problems in COVID-19 patients.Case presentation: We present two cases of patients with COVID-19-associated recurrent diarrhea and positive fecal occult blood who successfully recovered after a one-time convalescent plasma administration.Conclusion: When COVID-19 patients develop recurrent or refractory gastrointestinal symptoms and fail to respond to the available treatment, alternative therapy with convalescent plasma administration may be considered.
文摘Background: Telmisartan is a highly variable drug which is used to treat hypertension. This study compared to compare the bioavailability of two 40 mg Telmisartan tablets in adult and healthy Bangladeshi subjects. Materials and Method: This study was open label, randomized, laboratory blind, single dose, three periods, two treatments, three-sequence, partial-replicate, crossover and comparative oral bioavailability study. In this study, 18 Bangladeshi subjects were enrolled and 17 subjects were completed. Serial blood samples were collected up to 96 hours following drug administration. By using Liquid Chromatography Mass Spectrometry (LC-MS/MS) method, plasma concentrations of Telmisartan were determined. Results: The two formulations of Telmisartan were considered bioequivalent if 90% Confidence Interval (CI) fall within the range of 80.00% - 125.00% for AUC parameters and reference-scaled-average bioequivalence of 69.84% - 143.19% for C<sub>max</sub>. The 90% Confidence Interval for C<sub>max</sub>, AUC<sub>0-t</sub> & AUC<sub>0-∞</sub> was found 84.88% - 107.79%, 89.76% - 109.55%, and 91.20% - 114.52%, respectively. Conclusion: According to rate and extent of absorption with the single dose of Reference Product (R): Micardis<sup><sup><sup>®</sup></sup></sup> 40 mg Tablet, under fasting conditions, a single dose of Telmisartan 40 mg Tablet is bioequivalent.
文摘We present a new method of treatment for nasal valve dysfunction caused by insufficiency or stenosis in a patient who refused open septum revision despite a significant degree of septum deviation. The Anterior Spreader Flap (ASF) technique was suggested as an alternative to open nasal septum revision and was performed under local anesthesia. Computational fluid dynamics (CFD) tests were performed pre- and post-operatively and our patient was asked to complete a self-assessment using a Visual Analog Scale (VAS) for nasal breathing (0 = free nasal breathing, 10 = complete nasal blockage) before and 12 months after surgery. The ASF is a minimally invasive endonasal procedure in which the caudal edge of the upper lateral cartilage is dissected from the septum, and folded inwards and fixed. The ASF allows for less airflow resistance and more free space. CFD techniques revealed a reduction in local pressure based on extended space. In addition, VAS scores improved from 9 to 2 points (right side) and from 8 to 2 points (left side). On the basis of these findings, the ASF technique can be considered a safe, minimally invasive spreader flap technique. It can easily be combined with other nasal surgical techniques, as it is necessary in most cases. In selected cases, the ASF may be performed as a single procedure in patients with nasal valve dysfunction caused by septum deviation as an alternative to open septum revision.
文摘Objective To investigate the clinical application of Real-Time PCR for rapid detection of methicillin-resistant Staphylococcus aureus(MRSA) directly from nasopharyngeal swab specimens.Methods We collected the nasal and throat swab specimens from patients or medical staffs in 3 intensive care units,blood laminar flow ward and respiratory ward in Beijing Hospital,Ministry of Health from December 2010to April 2011.Each sample was tested by RT-PCR and conventional culture-based method for the presence of MRSA.Results The total number of the specimens was 206.Compared with the conventional culture-based method,we demonstrated the diagnostic values for Real-Time PCR were 96.4%sensitivity,96.6%specificity,81.8%positive predictive rate,and 99.4%negative predictive rate.And the limit of detection was 10~2CFU/ml.Conclusions This Real-Time PCR is a simple,rapid,sensitive and specific method.With the high negative predictive value,it can be used for the exclusion of MRSA colonization or infection.However,the application of its low positive predictive value should be further evaluated.
文摘Novel coronavirus,SARS-CoV-2 is responsible for causing a pandemic that has affected individuals worldwide,over 192 million people and about 4.1 million people died so far.The spread is ongoing and the numbers are still increasing.Numerous therapeutic approaches have been explored and developed during this pandemic.Immunotherapy with virus-specific antibodies in convalescent plasma(CP)has shown potential benefits for various pathogenic diseases.In many instances,it is the only available and safe management option for the COVID-19 patients.Here we describe two confirmed cases of COVID-19 from two different geographical areas that were managed with standard treatment modalities initially.Both of the patients were presented with high-grade fever,dry cough,and sore throat.Lab reports showed increased values of D-dimer,serum ferritin,leukocyte count(LC),Lactate dehydrogenase(LDH),and C-reactive protein(CRP).Chest X-ray showed bilateral infiltration(multifocal and bilateral ground-glass opacities and consolidations with peripheral and basal predominance),consistent with the previous reports on COVID-19 infection.The patients received conservative treatment according to the hospital's protocol.The convalescent plasma(from recovered patients)infusion was the last treatment given to both patients.After the convalescent plasma transfusion,both patients showed a reduction of viral load,an increase of anti-SARS-CoV-2 IgG and IgM antibodies,reduction in lung infiltration,with no adverse events.However,further randomized controlled trials are needed to investigate the full scope of safety and efficacy(both short and long-term)of convalescent plasma therapy for COVID-19 and other related infections.
基金supported by the National Natural Science Foundation of China (Nos.82072377 and 81971919)。
文摘Ecological studies suggested a link between air pollution and severe COVID-19 outcomes,while studies accounting for individual-level characteristics are limited.In the present study,we aimed to investigate the impact of short-term ambient air pollution exposure on disease severity among a cohort of 569 laboratory confirmed COVID-19 patients admitted to designated hospitals in Zhejiang province,China,from January 17 to March 3,2020,and elucidate the possible biological processes involved using transcriptomics.Compared with mild cases,severe cases had higher proportion of medical conditions as well as unfavorable results in most of the laboratory tests,and manifested higher air pollution exposure levels.Higher exposure to air pollutants was associated with increased risk of severe COVID-19 with odds ratio(OR)of 1.89(95%confidence interval(CI):1.01,3.53),2.35(95%CI:1.20,4.61),2.87(95%CI:1.68,4.91),and 2.01(95%CI:1.10,3.69)for PM_(2.5),PM_(10),NO_(2)and CO,respectively.OR for NO_(2)remained significant in two-pollutant models after adjusting for other pollutants.Transcriptional analysis showed 884 differentially expressed genes which mainly were enriched in virus clearance related biological processes between patients with high and low NO_(2)exposure levels,indicating that compromised immune response might be a potential underlying mechanistic pathway.These findings highlight the impact of shortterm air pollution exposure,particularly for NO_(2),on COVID-19 severity,and emphasize the significance in mitigating the COVID-19 burden of commitments to improve air quality.
