Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymph Nodes ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Vascular Endothelial Growth Factor C ; blood ; genetics ; metabolism

Administration, Oral ; Adult ; Atrial Flutter ; drug therapy ; physiopathology ; Digoxin ; therapeutic use ; Echocardiography ; Female ; Fetal Diseases ; drug therapy ; physiopathology ; Humans ; Hydrops Fetalis ; drug therapy ; physiopathology ; Infant, Newborn ; Male ; Pregnancy

A series of aralkyl-ketone-4-piperidol derivatives were synthesized and tested for their analgesic activities. All of the novel 30 compounds were prepared from 4-piperidone and alpha-halo-aralkyl-ketone through five steps, including Boc protection, nucleophilic addition in presence of CeCl3/NaI catalyst, deprotection, condensation and salification. Their structures were confirmed by 1H NMR and HRMS. Preliminary in vivo pharmacological trials showed that most of the synthesized compounds revealed analgesic effects. Among the tested compounds, 8, 13 and 22 exhibited potent analgesic activities in both mice writhing and mice hot plate model. The three compounds have low affinity for mu, delta, kappa receptors, which is a chance to find a better precursor of non-opioid analgesic for further optimization.
Analgesics, Non-Narcotic ; chemical synthesis ; chemistry ; pharmacology ; Animals ; Mice ; Molecular Structure ; Pain Measurement ; Pain Threshold ; drug effects ; Piperidones ; chemical synthesis ; chemistry ; pharmacology ; Receptors, Opioid, delta ; metabolism ; Receptors, Opioid, kappa ; metabolism ; Receptors, Opioid, mu ; metabolism ; Structure-Activity Relationship

OBJECTIVETo investigate the expressions of TopI gene in small cell lung cancer cell line H446, and explore the influence of TopI on the chemosensitivity of the cell line to topotecan (TPT).

METHODSWestern blot was performed to detect the TopI expression in H446 cells. Lipofectamine 2000 was used for the transient transfection of H446 cells by siRNA, and the transfection efficacy was detected. TopI mRNA was analyzed by quantitative RT-PCR and TopI protein was detected by Western blot to selected effective siRNA. The drug-sensitivity to topotecan (TPT) was evaluated by MTT assay.

RESULTSTopI gene was expressed in H446 cells. Lipofectamine 2000 mediated the siRNA effectively (88.67%). Compared with its parental cells, RT-PCR results showed that TopI mRNAs in transfected cells were reduced by (95.7 +/- 1.6)%, (90.8 +/- 1.6)%, (96.1 +/- 2.7)% and (96.3 +/- 1.8)%, respectively, and decreased significantly at protein level. By MTT assay, the inhibition rate of TPT to H446 cells transfected by siRNA was lower than that of control group at same concentrations (P < 0.01).

CONCLUSIONsiRNAs can silence the expression of TopI and decrease the drug-sensitivity of H446 cells to TPT.

Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Topoisomerases, Type I ; genetics ; metabolism ; Down-Regulation ; Drug Resistance, Neoplasm ; Humans ; Lung Neoplasms ; metabolism ; pathology ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Small Cell Lung Carcinoma ; metabolism ; pathology ; Topotecan ; pharmacology ; Transfection

【Objective】 To explore the clinical value of urinary aldosterone concentration(UAC) in primary aldosteronism(PA) screening and to evaluate the consistency of the two methods, liquid chromatography tandem mass spectrometry(LC-MS/MS) and chemiluminescence immunosorbent assay(CLIA), for determining UAC. 【Methods】 Among the 133 patients with suspected PA enrolled from October 2018 to August 2019, 55 were diagnosed with PA(30 with aldosteroneproducing adenoma and 25 with bilateral idiopathic hyperplasia) and 78 with essential hypertension(EH). Parallel determination of UAC was done with LC-MS/MS and CLIA assays on all subjects, then we compared the two methods and evaluated their correlation and consistency. Reciever operating characteristic(ROC) analysis was applied to assess the diagnostic accuracies, area under the curve(AUC) and cutoff values of UAC, plasma aldosterone concentration(PAC), random aldosterone to renin ratio(ARR) and urine aldosterone to renin ratio(UARR). Furthermore, ROC analysis of high urine sodium subgroup(urine sodium≥130 mmol/24h) was performed to evaluate the impact of sodium intake on PA screening. 【Results】 ①UACCLIA and UACLC-MS/MS were positively correlated(r=0.69, P < 0.001). Bland-Altman analysis showed poor consistency between CLIA and LC-MS/MS. ②The diagnostic efficiency of UAC and PAC in PA screening was equivalent (P > 0.05), while ARR was more efficient than UAC and PAC(P < 0.0001). AUC of UACCLIA, UACLC-MS/MS, PAC and ARR was 0.739, 0.659, 0.723, 0.943, respectively. The cutoff values for them were 5.1 μg/24 h, 11.6 μg/24 h, 27.6 ng/dL and 23.2 ng·dL^-1(/ μg·L^-1·h^-1), respectively. ③Both UARRCLIA and UARRLC-MS/MS performed as well as ARR in PA screening. AUC of UARRCLIA was 0.924 at the cutoff value of 1.75 μg·24h^-1(/ μg·L^-1·h^-1). While AUC of UARRLC-MS/MS was 0.906 at the cutoff value of 4.63 μg·24h^-1(/ μg·L^-1·h^-1). AUC of UACLC-MS/MS in high urine sodium subgroup was 0.708 at the cutoff of 14.9 μg/ 24 h. ④The diagnostic efficiency of UAC made no difference after sodium intake increased to 130 mmol/24 h or higher. 【Conclusion】 UAC alone is of little clinical value compared to ARR in PA screening and can serve as a supplementary indicator currently. But UARR, a combintion of UAC and plasma renin activity(PRA), is as valuable as ARR for screening PA. The optimal cutoff values of UACLC-MS/MS and UACCLIA are 11.6 μg/24 h, 5.1 μg/24 h, respectively. The optimal cutoff values of UARRLC-MS/MS and UARRCLIA are 4.63 μg·24 h^-1/(μg·L^-1·h^-1) and 1.75 μg·24 h^-1/(μg·L^-1·h^-1), respectively. Besides, significant bias of UAC observed in CLIA and LC-MS/MS results indicates that appropriate definition of cutoff value and reference range for each method are mandatory.

ObjectiveTo explore the value of plasma free metanephrines， including metanephrine（MN） and normetanephrine（NMN）， collectively referred to as MNs in the diagnosis of pheochromocytoma and paraganglioma （PPGL）. MethodsThe study included 1 631 patients suspected of PPGL from December 2014 to December 2020 in SunYat-sen Memorial Hospital. In these subjects， Plasma free MNs were measured by liquid chromatography-tandem mass spectrometry（LC-MS/MS） before surgery. Receiver operating characteristic （ROC） curves were used to determine the sensitivity and specificity of plasma NMN and MN in the diagnosis of PPGL. ResultsOf the screened patients， 108 patients had pathologically confirmed PPGL and 1 523 patients had definitive diagnoses other than PPGL as a control group. The median value of plasma MN ［0.54（0.17~4.48） nmol/L vs. 0.15（0.11~0.21） nmol/L， P<0.001］ and NMN ［7.48（2.12~15.01） nmol/L vs. 0.32（0.22~0.46） nmol/L， P<0.001］ were significant higher in the PPGL group than in the control group. Using an upper cut-off of 0.395 nmol/L for MN， the sensitivity was 60.2% and the specificity was 97.8%； when using a cut-off of 1.105 nmol/L for NMN， the diagnostic sensitivity was 87.0%， and the specificity was 98.7%. The area under the ROC curve and 95% confidence interval of plasma MN and NMN combined were 0.800（0.743， 0.858）， 0.959（0.932， 0.98） and 0.970（0.944， 0.996）， respectively. Analyzing the false-positive results， it was found that in the control group， 3% of the false-positive cases appeared. The estimated glomerular filtration rate（eGFR） were significantly lower in the false-positive group than in the true-negative group ［74.42（51.04~96.96） mL/min vs. 88.51（72.80~101.83） mL/min， P=0.001］. In 212 patients with eGFR lower than 60 mL/min， the false positive rate was increased to 8%. If the upper limit of the reference interval was increased by 25%， the specificity was increased to 96.7%； when the upper limit of the reference interval was increased by 50%， the specificity was 98.6 %. ConclusionsPlasma free MNs has a great reliability in the diagnosis of PPGL. The negative predictive value is extremely high and close to 100%. Combination analysis of plasma MN and NMN can further improve diagnostic performance. However， a few false positive cases may appear and might be influenced by impaired renal function. In patients with eGFR lower than 60 mL/min， the false positive rate is significantly increasing， which need a 25%-50% increase in the expected upper limit of a reference range to guarantee high specificity.

OBJECTIVETo study the anti-inflammatory mechanisms of total glycosides of Acanthopanax Giraldii (TGA).

METHODSThe changes of prostaglandin E(2)(PGE(2)), tumor necrosis factor (TNF-alpha), nitric oxide (NO), and expressions of COX-1 mRNA and COX-2 mRNA in BALB/c mouse macrophages were observed by the radioimmunoassay, ELISA and nitric acid reduction and RT-PCR in the presence or absence of TGA.

RESULTS(1) TGA could significantly decrease the production of PGE(2)and NO in mouse peritoneal macrophages. The inhibitory rate to LPS-induced PGE(2)production was 87% (TGA 100 mg/L, P<0.05, vs. LPS) and 62% (TGA 20 mg/L, P<0.05, vs. LPS), respectively. The inhibitory rate of NO production in mouse peritoneal macrophages was 49% (TGA 100 mg/L, P<0.05, vs. LPS) and 21% (TGA 20 mg/L, P<0.05 vs. LPS), respectively. TGA could not inhibit LPS-induced TNF-alpha production in mouse peritoneal macrophages. (2) TGA also inhibited the expression of COX-1 and COX-2 mRNA in RAW264.7 cells. The inhibitory rate of TGA to COX-1 mRNA was 22% (TGA 100 mg/L, P<0.05, vs. blank). The inhibitory rate of TGA to COX-2 mRNA was 55% (TGA 20 mg/L, P<0.05, vs. LPS) and 100% (TGA 100 mg/L, P<0.01 vs. LPS), respectively.

CONCLUSIONThe anti-inflammatory mechanisms of TGA for inhibiting the production of NO and PGE(2)are through inhibiting COX-2 mRNA expression without TNF-alpha changes.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Cell Line ; Cyclooxygenase 1 ; genetics ; Cyclooxygenase 2 ; genetics ; Dinoprostone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Eleutherococcus ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Glycosides ; pharmacology ; Lipopolysaccharides ; pharmacology ; Macrophages, Peritoneal ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred BALB C ; Nitric Oxide ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To discuss a real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) wether if can be used to detect Brucella. Methods According to the BCSP31 gene sequences specific for Brucella, one pair of primers and one TaqMan probe were designed. A real-time PCR was developed with the BCSP31 fragments cloned into PMD18-T vector. The standard cure was established and the sensitivity, the species specificity and the stability of the assay were evaluated. The clinical blood specimens were detected by QT-PCR and compared with clinical diagnosis. Results The standard curve was established with the standard template and the relationship between the Ct and the DNA copy number was linear(r=0.999). The sensitivity of the real-time PCR was 5 copies/μl. The sensitivity of the common PCR was 5×102 copies/μl. The sensitivity was about 100 times higher than common PCR. Species specificity of this FQ-PCR assay evaluated using genomic DNA from 6 Bmcella strains and 5 non-Brucella strains and strong fluorescence was detected in all Brucella strains. The CV of intra-assay and inter-assay reproducibility were 0.71%,7.23%, reprectively. Twenty-four specimens from clinical brucellosis cases, 19 showed positive, the positive coincident rate was 79%(19/24). The negative results were obtained for all 31 negative control, and the negative coincident rate was 100%(31/31). Two were positive from all 30 specimens clinically suspected. Conclusions Highly specific, sensitive, repeatable and coincidental with clinic, this FQ-PCR is quite useful for rapid detection of tiny DNA of Brucella in various samples and laboratory diagnosis.

OBJECTIVETo investigate the relationship between the expressions of integrin alpha5 beta1 and E-CD, and clinicopathological characteristics and prognosis of patients with non-small cell lung carcinoma (NSCLC).

METHODSThe expression of integrin alpha5 beta1 and E-CD were analyzed in 53 NSCLC and 12 control specimens by immunohistochemical assay.

RESULTSThe expression of integrin alpha5 beta1 was significantly higher in NSCLC (58.5%) than that in normal lung tissue (16.7%), and also positively related with pathological characteristics (P = 0.021), lymph node metastasis (P = 0.006), and clinical stage (P = 0.002). The 3-year survival rate in NSCLC group was significantly lower than that in control group (22.3% vs 40.6% , P = 0.041). The positive expression of E-CD in NSCLC and control group was 32.1% and 91.7%, respectively, and negatively correlated with pathological characteristics (P = 0.010) and lymph node metastasis (P = 0.002). The 3-year survival rate in control group was 19.9%, lower than that in NSCLC group (41.2%, P > 0.05), but the difference is not significant.

CONCLUSIONThe overexpression of integrin alpha5 beta1 may contribute to lymph node metastasis and play an inverse role, while E-CD may be a beneficial prognostic factor in patients with NSCLC.

Adult ; Aged ; Cadherins ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Integrin alpha5beta1 ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Smoking ; Survival Analysis

OBJECTIVETo investigate the effectiveness, safety and possible mechanism of recombinate human interleukin 11 (rhIL-11) in the treatment of chemotherapy-induced thrombocytopenia.

METHODSThirty-four patients (totally 76 cycles) with chemotherapy-induced thrombocytopenia received subcutaneous injection of rhIL-11 at the dose of 25 microg.kg(-1).d(-1) for 4 to 16 days. Serum IL-11 level was measured by ELISA, and IL-11 R alpha expression was detected by RT-PCR.

RESULTSThe mean baseline platelet count before chemotherapy was (135.0 +/- 54.3) x 10(9)/L for the 1st cycle and (259.4 +/- 64.5) x 10(9)/L for the 2nd cycle. The time to administer rhIL-11 was 7 to 16 days (median 12 days) in the 1st cycle and 4 to 10 days (median 6 days) in the 2nd, respectively (P < 0.05). The duration of post-chemotherapy platelet count below 50 x 10(9)/L was 7 to 13 days (median 10 days) for the 1st cycle and 3 to 8 days (median 5 days) for the 2nd, respectively (P < 0.05). Platelet count reached 300 x 10(9)/L or above in 30 chemotherapy cycles. The maximum platelet count was found to appear at D10 to D 17 (median D14), and negatively correlated with the pre-chemotherapy serum IL-11 level after administration of rhIL-11. Major adverse reactions included edema, headache, muscle and joint pain.

CONCLUSIONrhIL-11 is effective and safe for the treatment of chemotherapy-induced thrombocytopenia, with a relatively slow but sustained effect on the recovery of platelet count. Pre-chemotherpy serum IL-11 level might predict the efficacy of rhIL-11.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Breast Neoplasms ; drug therapy ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Female ; Humans ; Injections, Subcutaneous ; Interleukin-11 ; administration & dosage ; adverse effects ; blood ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Platelet Count ; Recombinant Proteins ; administration & dosage ; adverse effects ; Thrombocytopenia ; chemically induced ; drug therapy ; Treatment Outcome