Activin B signaling may promote the conversion of normal fibroblasts to scar fibroblasts.

This study is to explore the molecular mechanism of benign bile duct hypertrophic scar formation.Differential proteins between the normal fibroblast (NFB) and scar fibroblast (SCFB) were screened by protein chip assay, and analyzed by pathway-enrichment analysis and function-enrichment analysis. The differential proteins were further tested by ELISA. SiRNA-Act B was transfected to SCFB to down-regulate the expression of Act B. NFB was incubated with rh-Act B. The cell apoptosis and cell cycle were determined by flow cytometry. The expression of Act B, Smad2/3, transforming growth factor-ß1 (TGF-ß1), endothelin-1 (ET-1), thrombospondin-1 (Tsp-1), and Oncostatin M (OSM) were detected by Western blot.A total of 37 differential proteins were identified in SCFBs by microarray (P < .05), including 27 up-regulated proteins and 10 down-regulated proteins (P < .05). Their function were associated with Activin signaling, synthesis and degradation of extracellular matrix, formation and activation of cytokine, inflammatory reaction, immunoreaction, tissue damage reaction, cell cycle, migration, apoptosis, and secretion, etc. ELISA results showed that the expression of Act B, TGF-ß1, ET-1 were higher in SCFBs, while the expression of Tsp-1 and OSM were lower in SCFBs (P < .05). After interfered by siRNA-Act B, the expression of Act B mRNA decreased (P < .05). The percentage of early apoptosis increased (P < .05). The expression of Act B, Smad2/3, TGF-ß1 were decreased and Tsp-1, OSM were increased (P < .05). After treatment with rh-Act B, the percentage of G0/G1 phase of NFBs was decreased and that of S phase was increased without significance (P > .05). The expression of Act B, Smad2/3, TGF-ß1 were increased (P < .05) and Tsp-1, OSM were decreased (P < .01).There are differentially expressed proteins between SCFBs and NFBs. Activin B signal plays an important role in the process of NFB transforming to SCFB, and TGF-ß1, Smad2/3, Tsp-1, and OSM are important participants.

Combined use of NLR, V2o and Dmean to predict radiation-induced lung injury in lung cancer patients: an external validation study / 中华放射肿瘤学杂志

Objective To externally validate the accuracy of combined use of neutrophil-lymphocyte ratio (NLR),V20,and Dmean in predicting the incidence of grade Ⅲ or higher radiation-induced lung injury (RILI) in lung cancer patients.Methods A total of 166 lung cancer patients,who participated in the model establishment were selected into the internal validation group,and 85 lung cancer patients who received intensity-modulated radiotherapy in our department between June 2016 and June 2018 were assigned into the external validation group.The incidence rate of grade 3 or higher RILI was statistically compared between the internal and external validation groups.Multivariate logistic analysis was performed for NLR,V20 and Dmean The discrimination degree of the predictive model was evaluated by using ROC curve in combination with NLR,V20 and Dmean The calibration degree of the predictive model was assessed by Hosmer-Lemeshow test.Results The incidence rate of grade 3 or higher RILI in the internal and external validation groups was 23.8％ and 22.9％.Multivariate logistic analysis demonstrated that NLR,V20 and Dmean significantly differed in the internal validation group (P=0.032,0.006 and 0.005).However,only V20 significantly differed in the external validation group (P=0.038).The discrimination and calibration degree of RILI was almost consistent between the internal and external validation groups (both P＞0.05).The area under the curve (AUC) predicted by NLR,V20,Dmean and the combination of three indexes were 0.611,0.646,0.682 and 0.775 in the internal validation group,and 0.544,0.702,0.658 and 0.754 in the external validation group,respectively.The calibration degree in the internal validation group was P=2.797and 0.834,P=2.452 and 0.653 in the external validation group.Conclusion Combined application of NLR,V20 and Dmean can accurately predict the incidence of grade Ⅲ or higher RILI in lung can cancer patients,which has been validated by external dataset.

Qualitative and Quantitative Study of Tibetan Medicine Thlaspi semen / 中国药房

OBJECTIVE： To establish the qualitative and quantitative control method of Tibetan medicine Thlaspi semen. METHODS： TLC and HPLC method were used to identify and determine flavonoids isovitexin， swertisin and glucosinolates sinigrin from 15 batches of T. semen. The stationary phases identified by TLC of flavonoids and glucosinolates were polyamide film and high performance silica gel GF254. The developing agents were trichloromethane-methanol-glacial acetic acid （11 ∶ 1 ∶ 1，V/V/V） and ethyl acetate-methanol- triethylamine （4 ∶ 5 ∶ 0.5，V/V/V）. In chromatogram condition of content determination of isovitexin and swertisin， the separation was performed on CAPCELL PAK MGⅡ C18 column with mobile phase composed of acetonitrile-0.4％ glacial acetic acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 336 nm. In chromatogram condition of content determination of sinigrin， the separation was performed on CAPCELL PAK MGⅡ C18 column with mobile phase composed of acetonitrile-0.02 mol/L tetrabutylammonium hydrogen sulfate （15 ∶ 85，V/V，pH 6） at the flow rate of 1.0 mL/min. The detection wavelength was set at 227 nm. RESULTS： In TLC identification chromatogram， spots corresponding to isovitexin， swertisin and sinigrin control were detected in test samples. The linear ranges of isovitexin， swertisin and sinigrin were 1.26-79.00， 1.21-75.38， 12.80-640.00 μg/mL， respectively （all r≥0.999 5）. The limits of detection （LODs） were 0.09， 0.12， 0.15 μg/mL， and limits of quantitation （LOQs） were 0.39， 0.43， 0.54 μg/mL， respectively. RSDs of precision， stability （24 h） and reproducibility tests were all lower than 2.0％（n＝6）. The recoveries were 99.1％， 97.0％ and 98.1％， and RSDs were 1.9％， 1.8％， 1.8％（n＝6），respectively. The contents of isovitexin， swertisin and sinigrin in 15 batches of T. semen were 0.013-0.090， 0.020-0.130 and 18.92-40.75 mg/g， respectively. CONCLUSIONS： Established quality control method is simple， reproducible and stable， and can be used for the quality control of Tibetan medicine T. semen.

Dynamic change of fatigue of pemetrexed maintenance treatment in the JMEN trial.

OBJECTIVES: In the JMEN trial, patients with advanced non-squamous non-small cell lung cancer (NSCLC) without progression after platinum-based first-line therapy derived extended survival, delayed disease progression, and maintained overall quality of life (QoL) from pemetrexed maintenance therapy. However, fatigue was the most common physician-reported non-hematological toxicity in the pemetrexed group. This post hoc analysis investigated dynamic change of fatigue. MATERIALS AND METHODS: Analysis of the overall safety population with squamous and non-squamous NSCLC subgroups included Common Terminology Criteria for Adverse Events to summarize adverse event (AE) rates by cycle and AE investigator-reported severity. Worsening of fatigue, defined as +15mm or more from baseline on a 100mm scale, evaluated QoL using the patient-reported Lung Cancer Symptom Scale. Patients with worsening fatigue and time-to-worsening of fatigue symptoms were also analyzed. RESULTS: Drug-related fatigue occurred more frequently with pemetrexed than placebo. The drug-related grade 3/4 fatigue was also higher in the overall population on pemetrexed than with placebo. Fatigue incidence during pemetrexed maintenance after induction was not altered with cumulative exposure. Percentage of patients who experienced worsening of fatigue based on patient-reported LCSS scores was comparable between the two arms in cycles 1-10. The time-to-worsening of fatigue was similar between the pemetrexed arm and the placebo arm in the overall population; however, the East Asian subpopulation patients taking pemetrexed experienced a longer median time-to-worsening of fatigue than patients taking placebo. CONCLUSION: Analyses suggest that despite higher incidence of any grade drug-related fatigue compared with placebo in patients with advanced NSCLC, pemetrexed maintenance does not impair patient-reported QoL.

Comparative study of three different methods in the determination of length of gross target volume for esophageal cancer radiotherapy / 中华放射肿瘤学杂志

Objective Compared with chest CT,endoscopic ultrasonography (EUS) can more accurately determine the upper and lower margins of esophageal cancer,and marking the upper and lower margins of the esophageal cancer with titanium clip contributes to the delineation of target area of esophageal cancer during radiotherapy.To compare the effects of esophageal X-ray,chest computed tomography (CT)scan and EUS-assisted placement of marker clip in the determination of the length of gross target volume (GTV),aiming to provide reference for the determination of GTV during esophageal cancer radiotherapy.Methods Thirty patients who were initially diagnosed with thoracic esophageal cancer by histological and cytological examinations and scheduled to receive radiotherapy were recruited in this investigation.All patients received esophageal X-ray,CT scan,and EUS-assisted placement of marker clip.The length of GTV was quantitatively measured and statistically compared among three different methods.Results The length of GTV was (6.1 ± 1.4) cm,(6.8± 1.9) cm and (6.3± 1.9) cm determined by esophageal X-ray,CT scan and EUS-assisted placement of marker clip,respectively.Compared with CT scan,the length of GTV determined by EUS-assisted placement of marker clip did not significantly differ (P=0.11).The length of GTV determined by esophageal X-ray was significantly shorter than that by CT scan (P =0.03).Among all patients,the length of GTV determined by EUS-assisted placement of marker clip was longer compared with that by chest CT scan in 22.2％ of patients.The length of GTV determined by EUS-assisted placement of marker clip was the same as that by chest CT scan in 11.1％ of patients.The length of GTV determined by EUS-assisted placement of marker clip was shorter compared with that by chest CT scan in 66.7％ of patients.Conclusions EUS-assisted placement of marker clip differs from esophageal X-ray and CT scan in determining the length of GTV,which acts as one of the effective methods in the determination of the length of GTV during esophageal cancer radiotherapy.

Study on Quality Standard for Tibetan Medicine Jiuwei Zhaxun Pills / 中国药师

Objective: To establish the quality standard for Jiuwei Zhaxun pills. Methods: Fructus Chebulae, Flos Carthami and Herba Dracocephali Heterophylli were identified by TLC. HPLC was used to determine the content of gallic acid in Jiuwei Zhaxun pills. Results: TLC showed clear sports without any interference from the negative control. Gallic acid showed a good linear relationship (r=0. 999 7) within the range of 0. 028 6-0. 572 0 μg. The average recovery was 97. 74% with the RSD of 0. 95% (n=6). Conclusion:The methods are with strong specificity, high sensitivity and good reproducibility, which can be applied in the quality control of Jiuwei Zhaxun pills.

A multicenter survey of first-line treatment patterns and gene aberration test status of patients with unresectable Stage IIIB/IV nonsquamous non-small cell lung cancer in China (CTONG 1506).

BACKGROUND: In recent years, systemic chemotherapy and molecular targeted therapy have become standard first-line treatments for locally advanced or metastatic nonsquamous non-small cell lung cancer (NSCLC). The objective of this survey was to investigate first-line anticancer treatment patterns and gene aberration test status of patients with advanced nonsquamous NSCLC in China. METHODS: Patients included in this study had unresectable Stage IIIB/IV nonsquamous NSCLC and were admitted during August 2015 to March 2016 into one of 12 tertiary hospitals throughout China for first-line anticancer treatment. Patient data (demographics, NSCLC histologic type, Eastern Cooperative Oncology Group [ECOG] Performance Status [PS], gene aberration test and results [if performed], and first-line anticancer treatment regimen) were extracted from medical charts and entered into Medical Record Abstraction Forms (MERAFs), which were collated for analysis. RESULTS: Overall, 1041 MERAFs were collected and data from 932 MERAFs were included for analysis. Patients with unresectable Stage IIIB/IV nonsquamous NSCLC had a median age of 59 years, 56.4% were male, 58.2% were never smokers, 95.0% had adenocarcinoma, and 92.9% had an ECOG PS ≤1. A total of 665 (71.4%) patients had gene aberration tests; 46.5% (309/665) had epidermal growth factor receptor (EGFR) gene mutations, 11.5% (48/416) had anaplastic lymphoma kinase (ALK) gene fusions, and 0.8% (1/128) had a c-ros oncogene 1 gene fusion. The most common first-line treatment regimen for unresectable Stage IIIB/IV nonsquamous NSCLC was chemotherapy (72.5%, 676/932), followed by tyrosine kinase inhibitors (TKIs; 26.1%, 243/932), and TKIs plus chemotherapy (1.4%, 13/932). Most chemotherapy regimens were platinum-doublet regimens (93.5%, 631/676) and pemetrexed was the most common nonplatinum chemotherapy-backbone agent (70.2%, 443/631) in platinum-doublet regimens. Most EGFR mutation-positive patients (66.3%, 205/309) were treated with EGFR-TKIs. CONCLUSIONS: Findings from our survey of 12 tertiary hospitals throughout China showed an increased rate of gene aberration testing, compared with those rates reported in previous surveys, for patients with advanced nonsquamous NSCLC. In addition, pemetrexed/platinum-doublet chemotherapy was the predominant first-line chemotherapy regimen for this population. Most patients were treated based on their gene aberration test status and results.

Relationship between catecholamine level and gene polymorphism of β1 adrenergic receptor G1165C in children with EV71 infection in hand foot and mouth disease

OBJECTIVE@#To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD).@*METHODS@#The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA).@*RESULTS@#The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P  0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05).@*CONCLUSIONS@#As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.

Relationship between catecholamine level and gene polymorphism of β1 adrenergic receptor G1165C in children with EV71 infection in hand foot and mouth disease

Objective To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD). Methods The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA). Results The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P 0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05). Conclusions As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.

SFRP5 gene silencing promotes proliferation and migration of human pancreatic cancer cell line PANC-1 / 基础医学与临床

Objective To investigate the effect of lentivirus-mediated shRNA silencing SFRP5 on proliferation,in-vasion and migration of pancreatic cancer cell line PANC-1. Methods A SFRP5-knockdown recombinant plasmid was constructed and successfully transfected it into pancreatic cancer cell line PANC-1,blank plasmid transfection was treated as negative control and untreated cells as blank control group. The expression of SFRP5 at RNA and protein level in cell were detected by real-time PCR and Western blot, CCK-8 assay was applied to examine the effect of SFRP5 silencing on the proliferation, the cell migration of pancreatic cancer cell line PANC-1 was ana-lyzed by Transwell migration assay and cell scratch test was used to examine the cell invasion in PANC-1 cell. Results Stable transfected shRNA-SFRP5 cell of pancreatic cancer line was established successfully.The prolifera-tion capacity of SFRP5 group was significantly higher as compared to the negative control and blank control group by CCK8 assay(P<0.01).Similarly, cell invasion and migration of SFRP5 group were significantly higher compared to the negative control and blank control group(P<0.01). Conclusions SFRP5 lentiviral interference vectors can effectively decrease SFRP5 gene expression in PANC-1 cell of pancreatic cancer, thereby promoting cell proliferation,invasion and migration.

SFRP5 gene silencing promotes proliferation and migration of human pancreatic cancer cell line PANC-1 / 基础医学与临床

Objective To investigate the effect of lentivirus-mediated shRNA silencing SFRP5 on proliferation,in-vasion and migration of pancreatic cancer cell line PANC-1. Methods A SFRP5-knockdown recombinant plasmid was constructed and successfully transfected it into pancreatic cancer cell line PANC-1,blank plasmid transfection was treated as negative control and untreated cells as blank control group. The expression of SFRP5 at RNA and protein level in cell were detected by real-time PCR and Western blot, CCK-8 assay was applied to examine the effect of SFRP5 silencing on the proliferation, the cell migration of pancreatic cancer cell line PANC-1 was ana-lyzed by Transwell migration assay and cell scratch test was used to examine the cell invasion in PANC-1 cell. Results Stable transfected shRNA-SFRP5 cell of pancreatic cancer line was established successfully.The prolifera-tion capacity of SFRP5 group was significantly higher as compared to the negative control and blank control group by CCK8 assay(P<0.01).Similarly, cell invasion and migration of SFRP5 group were significantly higher compared to the negative control and blank control group(P<0.01). Conclusions SFRP5 lentiviral interference vectors can effectively decrease SFRP5 gene expression in PANC-1 cell of pancreatic cancer, thereby promoting cell proliferation,invasion and migration.

Meta-Analysis of First-Line Pemetrexed Plus Platinum Treatment in Compared to Other Platinum-Based Doublet Regimens in Elderly East Asian Patients With Advanced Nonsquamous Non-Small-Cell Lung Cancer.

BACKGROUND: Pemetrexed plus platinum has become a standard of care in first-line treatment for patients with advanced nonsquamous non-small-cell lung cancer. However, elderly lung cancer patients are generally understudied and undertreated in clinical practice in East Asia because of safety concerns. This analysis aimed to provide a picture of the clinical benefit of pemetrexed/platinum in the first-line setting for elderly (age ≥ 65 years) East Asian patients. PATIENTS AND METHODS: Individual patient data from 3 randomized controlled phase 3 trials that enrolled East Asian patients were analyzed in this meta-analysis. RESULTS: In elderly East Asian patients (63 in the pemetrexed/platinum group and 42 in the control group), pemetrexed/platinum treatment achieved more benefits compared to other platinum-based doublets, including better overall response rate (32.8% vs. 7.5%), favorable progression-free survival (not statistically significant in adjusted hazard ratio), and significantly longer (3.15 vs. 1.54 months) survival without drug-related grade 3/4 toxicity. Overall survival was numerically prolonged (16.33 vs. 13.77 months; not statistically significant). These benefit trends were similar to those in all-age East Asian patients. In elderly East Asians, pemetrexed/platinum treatment was also associated with a lower incidence rate of drug-related grade 3/4 adverse events. The adverse event profile was similar to that in all-age East Asians. There were no unexpected adverse events. CONCLUSION: Pemetrexed/platinum had good efficacy and also resulted in better overall response and tolerability than other platinum-based doublets as first-line treatment in nonsquamous non-small cell lung cancer in elderly East Asians, which was consistent with data observed in all-age East Asians.

Chemical constituents from roots of Lindera aggregate / 中草药

Objective: To study the chemical constituents from the roots of Lindera aggregata. Methods: The compounds were isolated and purified by various column chromatographies. Their structures were elucidated by means of spectral analyses (MS, 1D-NMR, and 2D-NMR). Results: Fourteen compounds were isolated from 95% ethanol extract of L. aggregate. Compound 1 was isolated as a new one and named as bi-linderachalcone (1), and the other compounds were identified as methyllinderone (2), linderone (3), pashanone (4), pinostrobin (5), methyllucidone (6), pinocembrin (7), 5,7-dihydroxy-6,8-dimethoxyflavone (8), lucidone (9), ethyllucidone (10), cinnamic acid (11), pinostrobinchalcone (12), cirsimaritin (13), and β-sitosterol (14), respectively. Conclusion: Compound 1 is isolated as a new compound and compound 13 is firstly obtained from the plants of Lindera Thunb..

Relationship of expressions of HIF-1α, VEGF and EGFR in esophageal squamous cell carcinoma to radiation therapy efficacy / 实用医学杂志

Objective To probe into the relation of expressions of HIF-1α, VEGF and EGFR in esophageal squamous cell carcinoma with radiation therapy efficacy. Methods 73 of the patients with carcinoma of oesophagus from January , 2011 to May , 2014 in the General Hospital of Ningxia Medical University , were involved in this research , their clinical data reviewed and analyzed. Before radiotherapy , immunohistochemical SP was used to test expressions of HIF-1α, VEGF and EGFR in the cancer tissues. Relationships between the expressions and the efficiency of radiotherapy were analyzed. Results The positive expressions of HIF-1α, VEGF and EGFR were 70.0%, 84.9% and 80.8%, respectively. In terms of the single factor analysis related to recent curative effects, HIF-1α expression had significant correlation with recent curative effects (P=0.03). Conversely , multiplicity indicated that HIF-1α and EGFR expressions were notably associated with recent curative effects (P=0.007, 0.045, respectively). Conclusions The positive expressions of HIF-1α,VEGF and EGFR in the esophageal carcinoma may account for a largest proportion of the total. HIF-1α and EGFR expressions are associated with the short-term outcomes.

Ghrelin, neuropeptide Y (NPY) and cholecystokinin (CCK) in blunt snout bream (Megalobrama amblycephala): cDNA cloning, tissue distribution and mRNA expression changes responding to fasting and refeeding.

Blunt snout bream (Megalobrama amblycephala Yih, 1955) is an endemic freshwater fish in China for which the endocrine mechanism of regulation of feeding has never been examined. Ghrelin, neuropeptide Y (NPY) and cholecystokinin (CCK) play important roles in the regulation of fish feeding. In this study, full-length cDNAs of ghrelin, NPY and CCK were cloned and analyzed from blunt snout bream. Both the ghrelin and NPY genes of blunt snout bream had the same amino acid sequences as grass carp, and CCK also shared considerable similarity with that of grass carp. The three genes were expressed in a wide range of adult tissues, with the highest expression levels of ghrelin in the hindgut, NPY in the hypothalamus and CCK in the pituitary, respectively. Starvation challenge experiments showed that the expression levels of ghrelin and NPY mRNA increased in brain and intestine after starvation, and the expression levels of CCK decreased after starvation. Refeeding could bring the expression levels of the three genes back to the control levels. These results indicated that the feeding behavior of blunt snout bream was regulated by the potential correlative actions of ghrelin, NPY and CCK, which contributed to the defense against starvation. This study will further our understanding of the function of ghrelin, NPY and CCK and the molecular mechanism of feeding regulation in teleosts.

Chemical constituents of Oxytropis myriophy / 中草药

Objective: To study the chemical constituents of CHCl3 and EtOAc extracts from Oxytropis myriophy. Methods: The CHCl3 and EtOAc extracts of O. myriophy were isolated and purified by silica gel chromatography, Sephadex LH-20, and PTLC chromatography. The structures were identified by spectral methods. Results: Eleven compounds were obtained and identified from the CHCl3 and EtOAc extracts from O. myriophy, namely 5-hydroxyl-7,4'-dimethoxyflavone (1), 4-hydroxylacetophenone (2), 5,4'-dihydroxyl-7,3'-dimethoxyflavone (3), 5,7-dihydroxyl-6,4'-dimethoxyflavone (4), isorhamnetin (5), kaempferol (6), acetophenone- 4-O-β-D-glucoside (7), 2-hydroxyl-6-methoxyacetophenone-4-O-β-D-glucoside (8), 6-methoxycoumarin-7-O-β-D-glucoside (9), isorhamnetin-3-O-β-D-glucoside (10), and kaempferol-3-O-β-D-glucoside (11). Conclusion: All of the compounds are isolated from this plant for the first time and compound 1, 7, and 8 are isolated from the plants of Oxytropis DC. for the first time.

Molecular characterization of cholecystokinin in grass carp (Ctenopharyngodon idellus): cloning, localization, developmental profile, and effect of fasting and refeeding on expression in the brain and intestine.

Cholecystokinin (CCK) is a multi-functional brain-gut peptide in fish and mammals. To investigate the role of CCK in appetite regulation in fish, a 770-bp full-length cDNA sequence of CCK gene was obtained by RT-PCR and rapid amplification of cDNA ends methods in grass carp Ctenopharyngodon idellus. Homology analysis showed that the CCK cDNA sequence of grass carp had the highest similarity (90 %) to that of goldfish Carassius auratus and a higher similarity (>70 %) to those of other teleosts than to mammals. The PCR amplification using genomic DNA identified that the CCK gene of grass carp was comprised of three exons and two introns. Real-time quantitative PCR was used to detect CCK mRNA expression in adult tissues. High levels of gene expression were found in the hypothalamus and pituitary; moderate levels in the intestine, muscle and white adipose tissue; and low levels in other tissues. During early development (i.e., fertilized eggs to 35-day post-hatching larvae) the levels of CCK mRNA expression were higher during embryonic developmental stages than during post-hatch larval stages. Fasting decreased CCK mRNA expression levels in the brain and intestine, whereas refeeding resulted in an increase of expression. The results suggest that CCK mRNA expression has obvious tissue specificity and may have a role in feed intake regulation in grass carp.

[Human CD96 gene cloning, expression and identification].

OBJECTIVE: To construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om. METHODS: hCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting. RESULTS: hCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 kD hCD96om fusion protein. CONCLUSION: The CD96 gene of human has been successfully cloned and expressed in BL21(DE3) cells, and its rabbit polyclonal antibody has been obtained.

Molecular cloning and expression of orange-spotted grouper (Epinephelus coioides) CD8α and CD8ß genes.

T-cell surface glycoprotein CD8 consists of two distinguished chains, termed α and ß chains, and functions as a co-receptor for the T-cell receptor by binding to MHC class I proteins. In this study we report the cloning and identification of both CD8α and CD8ß genes from orange-spotted grouper (Epinephelus coioides). The predicted grouper CD8α and CD8ß proteins were structurally similar to other fish especially to those of Pleuronectiformes. Real-time RT-PCR revealed that the CD8 mRNA was much higher in the thymus than in other immune organs, and the expression level were very low in stomach, liver, and brain. During embryonic development of the grouper, the highest CD8 transcripts were detected in the multi-cell stage, followed by muscle burl stage, which suggested that the multi-cell stage may be critical in CD8 transcript synthesis. Moreover, CD8 mRNA levels were examined in lymphocytes at different time treated with lipopolysaccharide (LPS), polyriboinosinic polyribocytidylic acid (PolyI:C), phytohemagglutinin (PHA), and concanavalin A (ConA). The result showed that the CD8 mRNA levels were significantly affected in time-dependent manner by PolyI:C, PHA, and ConA, but not by LPS.

Human CD96 gene cloning, expression and identification / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om.</p><p><b>METHODS</b>hCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting.</p><p><b>RESULTS</b>hCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 kD hCD96om fusion protein.</p><p><b>CONCLUSION</b>The CD96 gene of human has been successfully cloned and expressed in BL21(DE3) cells, and its rabbit polyclonal antibody has been obtained.</p>