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.

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.

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.

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.

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.

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.

Determinaton of five flavonoids from different processing products of Artemisia frigida by HPCE / 中草药

Objective: Determining the content of five flavonoids in Artemisia frigida by high performance capillary electrophoresis (HPCE) method to provide reference data for the evaluation of processing products of A. frigida. Methods: Treated by different processing technologies, the content of five flavonoids compounds can be determined and compared by HPCE. Results: Regression equations were 5,7,3′-triterhydroxy-4′-methoxyflavone Y = 0.114 3 X + 0.032 7, r = 0.999 8; 5,3′-dihydroxy-6,7,4′-tritermethoxyflavone Y = 0.100 9 X + 0.048 5, r = 0.999 6; quercetin Y = 0.055 5 X + 0.026 8, r = 0.999 6; 5,7,3′-triterhydroxy-6,4′-dimethoxyflavone Y= 0.113 2X-0.016 1, r = 0.999 3; luteolin Y = 0.097 9X-0.029 5, r = 0.999 4. Five kinds of flavonoids were good linear relationship at 1.00-40.00, 10.00-200.00, 5.00-100.00, 1.00-40.00, 1.00-40.00 μg/mL. The average recoveries were 98.44%, 97.75%, 97.73%, 97.98%, and 98.07%. And RSD were 1.60%, 1.03%, 1.57%, 0.94%, and 1.20%. Conclusion: The results reveal that the different processing technologies have different effects on the content determination of five flavonoids compounds in A. frigida. Using the five flavonoids as testing indexes, the best processing technology of A. frigida is heating and drying to constant weight at 60 °C.

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>

Effects of Tangut Dragonhead (Dracocephalum tanguticum)on Blood Rheology and Tissuse Morphology in Rats / 中草药

The effects of an aqueous extract of Dracocephalum tanguticum Maxim. (DtM ) on p, b.b-1/ H. Hct. ESR. ESR-kand EPT of rats were studied by ip route. The animals were divided into three groups: normal pressure control group (NCG ). iowpressure treatment group (LTG )and lowpressure control group (LCG ). A11 p,b. Het. ESR-k increased stignificantly in rats exposed to stimulated altitude of 6500m for 10 days (8h/day),and the platelet and the ratio of left to ri1ght ventricle weights were obviously decreased in comparison with NPG. The above parameters of LTG .except Het.have noobvious difference as compared with NCG, Het in LTG increased obviously but lower than LCG. These results suggested that DtM may be used as an inhibitory agent against the changes of induced-hypoxia blood rheology, decreased platelet and hypertrophic right ventricle. In addition,the observation of tissue morphology showed that DtM possesses sometherapeutic effects to injuries of lung. liver and kidney of hypoxia rats.