1.Screening of differentially expressed genes in placentas with hepatitis B virus infection by suppression subtractive hybridization technique
Gui-Qin BAI ; Ya-Fei YUE ; Shu-Lin ZHANG ; Jun CHENG ; Yan LIU ; Shu-Hong LI ; Xin-E ZHANG ;
Chinese Journal of Obstetrics and Gynecology 2001;0(02):-
Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.
2.Preparation process of rutacarpine-hydroxypropyl-beta-cyclodextrin inclusion complex.
Chun-Lin YAN ; Ji ZHANG ; Yong HOU ; Gui-Ping XUE ; Shu WANG ; Qing-Ya ZHAO
China Journal of Chinese Materia Medica 2014;39(5):828-832
Rutaecarpine (Rut) is a type of indole quinazoline alkaloid exracted from Ruticarpum. Studies showed that Rut has a wide range of pharmacological effects, such as anti-hypertension, anticancer, anti-inflammation, anti-thrombus formation. Currently, many scholars are committed to developing it into a new antihypertensive and anti-inflammatory drug with all new mechanisms. But studies found that Rut is a highly fat-soluble drug with low water and oil solubility. Its high insolubility is the main obstacle in its oral absorption and application, which greatly reduced its bioavailability. Therefore, hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was used as the inclusion material to prepare Rut-HP-beta-CD inclusion complex in this experiment, in order to increase its water solubility and bioavailability. In this experiment, the inclusion complex was prepared by the stirring-freeze-dry method. The preparation process was optimized by the orthogonal test, with the inclusion rate as the index, and molar ratio between host and guest molecules, inclusion temperature, time and stirring speed as the impacting factors. Moreover, the inclusion complex was verified by detecting the apparent solubility, thin layer chromatography, microscopic identification, melting point detection and dissolution study. The results showed that under the conditions of the molar ratio between Rut and HP-beta-CD of 1: 1, temperature at 60 degrees C, inclusion time of 4h and stirring speed at 600 r x min(-1), the inclusion rate of Rut-HP-beta-CD reached 91.04%. Therefore, the preparation process of Rut-HP-beta-CD inclusion under the optimum conditions is simple and feasible, with a highest inclusion rate and reproducibility, and could significantly improve Rut's solubility and bioavailability, and provide a reliable experimental basis for its clinical application.
2-Hydroxypropyl-beta-cyclodextrin
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Alkaloids
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chemistry
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Chemistry, Pharmaceutical
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methods
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Drug Carriers
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chemistry
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Drugs, Chinese Herbal
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chemistry
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Rutaceae
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chemistry
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Solubility
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beta-Cyclodextrins
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chemistry
4.Reflections on the calculation methods of the imbalance degree in the biophysical properties of meridian points
Shu-Ya MA ; Li-Jia PAN ; Jun LIU ; Yan-Fen SHE
Journal of Acupuncture and Tuina Science 2021;19(4):271-277
Objective: To analyze and compare the features and strengths of different methods for calculating the biophysical properties of meridian points, and thus propose corresponding suggestions to fully achieve the research and application value of biophysical properties of meridian points.Methods: We searched and collected the literature on the imbalance of biophysical properties of meridian points between January 1, 2005 and March 1, 2020 in China National Knowledge Infrastructure (CNKI), Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP) and PubMed database, and then analyzed, compared, and summarized the applied methods for calculating the imbalance degree of the biophysical properties of meridian points. Results: The current methods for calculating the imbalance degree of the biophysical properties of meridian points are diverse and can be summarized as the following three: direct comparison of the measured values of the left and right namesake points, difference method, and ratio method. The low uniformity of the calculation methods has limited the promotion and application of its research results. Conclusion: In future research on the biophysical properties of meridian points, multidisciplinary cooperation in terms of imbalance degree calculation methods, detection instruments, and health data models is necessary to achieve more widely applicable scientific conclusions and more generalized experimental results.
5.Chemical kinetics study and application of a method with low usage amount of arsenic trioxide for determining urinary iodine by arsenite-ceric catalytic spectrophotometry using ammonium persulfate digestion
Ya-ping, ZHANG ; Yan-hong, HUANG ; Na, LI ; Shu-qiong, ZHANG
Chinese Journal of Endemiology 2013;(3):318-324
Objective To study the chemical kinetics characteristics in a new revised method with low usage amount of arsenic trioxide for determining urinary iodine by arsenite-ceric catalytic spectrophotometry using ammonium persulfate digestion,and to study the impact of operating bias in arsenite-ceric reaction temperature and reaction time on final results in this method.Methods The absorbances (A) of arsenite-ceric reaction of iodine standard series were measured at different reaction temperature and time,and the results were analyzed according to the chemical kinetics equation.The change values and half-life of A values of the new revised method and the current standard method were calculated.The chemical kinetics model of reaction system for this new revised method was deduced from experimental results.The calculation formula of result relative error for urinary iodine determination was deduced based on constants reaction temperature and reaction time and reaction rate constant factor.The result relative errors caused by operation deviation of reaction temperature or reaction time in the determination of urinary iodine were calculated.Results The usage amount of arsenious acid solution in the new revised method was only a quarter of usage amount of the current standard method(WS/T 107-2006).A values of each concentration of standard curve series at different reaction time t were obtained,the lnA to t mapping was a straight line,the linear correlation coefficients were-0.9995--0.9999.These results were in accord with the characteristic of chemical first-order reaction.Relationships between the reaction rate constant K and the reaction temperature T in the temperature range of 20-35 ℃ were well accord with Arrhenius equation.The A values and iodine concentrations (C) at various experimental temperatures showed good C =a + blnA linear relation,the absolute value of the linear correlation coefficient(| r |) > 0.9990.After calculation and comparison of changes in the half-life of A values in the new revised method and in the original standard method at 20,25,30,35 ℃ reaction temperature,half-life of A values of 0-300 μg/L iodine standard series in the new revised method and in the original standard method were 191.0-11.4 min and 66.8-10.2 min at 25℃,respectively.Under the same conditions of 25 ℃ for 40 min,the gradient of A values of 0-300 μg/L iodine standard curve in the new revised method was similar to that of the original standard method(slope-133.7,-139.2,respectively).But differences between A values of standard curve and the reaction initial absorbance(A0) in the original standard method were 1.4 to 3.7 times those of the new revised method.A chemical kinetics model of reacting system for this method was established.A series of urinary iodine results relative error data were obtained when reaction temperature deviation was ± 1,± 0.5,± 0.3 ℃ or reaction time deviation was ± 1 min for sample test tubes.Data showed that relative errors of urinary iodine results caused by reaction temperature deviation or reaction time deviation in the new revised method were less than those of the original standard method.Conclusions The iodine-catalyzed arsenite-ceric reaction in the new revised method is a first-order reaction,when measuring 0-300 μg/L urinary iodine at 20-35 ℃,and 300-1200 μg/L urinary iodine at 20-30 ℃,the calibration relation of C =a + blnA is established when arsenite-ceric catalytic spectrophotometry is kept at a certain stable temperature and in certain stable reaction time.Compared with the original standard method,using the revised method with low usage amount of arsenic trioxide for measuring urinary iodine,the arsenite-ceric reaction rate is slow down.As a result,this method is easier to operate and has better precision and accuracy.
6.Multivariate and multilevel model analysis on factors that influencing the literacy of health among high school students in Guangdong province
Xiao-Hua YE ; Ya XU ; Shu-Dong ZHOU ; Yan-Hui GAO ; Yan-Fen LI
Chinese Journal of Epidemiology 2011;32(9):873-876
Objective To analyze the awareness on health among high school students and its influencing factors in Guangdong. Methods Multi-stage sampling and questionnairs "2009 health awareness survey of the Chinese citizens" developed by our Department of Health, were used. Data were analyzed by multivariate multilevel model under MLwinN 2.19 software. Results The mean scores on knowledge and ideas, behaviors and related skills among 1606 high school students of Guangdong province, were 69.08 ± 14.81,60.05 ± 16.85 and 74.99 ± 21.17 respectively. Three items on health showed that they all related to each other and relations between grades (0.972,0.715 and 0.855) were greater than the individuals (0.565,0.426 and 0.438). Conclusion Factors as students from outside the Pearl River Delta region or from the rural areas, being male, at general secondary schools, at grade one, with poor academic performance and more pocket money etc., had lower levels on those related information of health.
7.Detection of W462X Mutation in Low Density Lipoprotein Receptor Gene of A Familial Hypercholesterolemia Patient and Its Clinical Significance
shu, LIU ; lu-ya, WANG ; jie, LIN ; qiang, YONG ; ya, YANG ; bang-jun, WU ; xiao-dong, PAN ; lan-ping, DU ; yan-wen, QIN
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To explore the molecular basis of familial hypercholesteraemia(FH)by analyzing the phenotype and genotype relationship through identify the low density liporotein receptor(LDL-r)gene mutation in a FH kindred.Methods A male patient of 15 years old was selected to examine the electrocardiogram,lipid.Color Doppler was used to examine heart and great vessels.The promoter region and the 18 exons of the LDL-r gene were screened by touch-down polymerase chain reaction(PCR)and DNA sequencing.Results The caro-tid intima-media thickness(IMT)was increased to 0.23 cm,while coronary flow velocity reserve(CFVR)was decreased to 1.57,and mode-rate mitral regurgitation was found in the proband.The genetic alteration G→A change at 1 448 of exon 10 causing premature stop codon(W462X).The same heterozygous nonsense mutation was also found in his father.The mutation had been reported in other Chinese patients.In vitro experiments showed that W462X mutation leads to low LDL binding and internalization ability.Conclusions The homozygous mutation(W462X)in exon 10 of the LDL-r gene were identified in the clinically heterozygous FH proband.The W462X mutation is the underl-ying cause of hypercholesterolaemia and clinical AS manifestations.W462X is recurrent mutation among Chinese FH patients.It might be a hot spot mutation in LDL-r in Chinese FH.J Appl Clin Pediatr,2009,24(1):18-20
8.Study on material base of Carthamus tinctorius with antioxidant effect based on selective knock-out.
Lin-Yan WANG ; Yu-Ping TANG ; Xin LIU ; Ya-Hui GE ; Shu-Jiao LI ; Er-Xin SHANG ; Jin-Ao DUAN
China Journal of Chinese Materia Medica 2014;39(7):1285-1289
OBJECTIVETo establish a method for studying efficacious materials of traditional Chinese medicines from an overall perspective.
METHODCarthamus tinctorius was taken the example. Its major components were depleted by preparing liquid chromatography. Afterwards, the samples with major components depleted were evaluated for their antioxidant effect, so as to compare and analyze the major efficacious materials of C. tinctorius with antioxidant activity and the contributions.
RESULTSeven major components were depleted from C. tinctorius samples, and six of them were identified with MS data and control comparison. After all of the samples including depleted materials are compared and evaluated for their antioxidant effect, the findings showed that hydroxysafflor yellow A, anhydrosafflor yellow B and 6-hydroxykaempferol-3, 6-di-O-glucoside-7-O-glucuronide were the major efficacious materials.
CONCLUSIONThis study explored a novel and effective method for studying efficacious materials of traditional Chinese medicines. Through this method, we could explain the direct and indirect contributions of different components to the efficacy of traditional Chinese medicines, and make the efficacious material expression of traditional Chinese medicines clearer.
Alkalies ; chemistry ; Antioxidants ; chemistry ; Carthamus tinctorius ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Mass Spectrometry
9.A method used in myeloma cell cloning spot image segmentation.
Yu-ya YAO ; Zhuang-zhi YAN ; Yu CHEN ; Shu-peng LIU
Chinese Journal of Medical Instrumentation 2008;32(5):319-322
A new interactive image segmentation method used in the multiple myeloma cloning spots image segmentation is presented in the paper. Based on the theory of graph cuts, some pixels are selected as the front object and the background seeds, and the other parts are treated as the unknown region. Then, an energy function is constructed and initialized through K-means, and the minimum cut method is used in the segmentation by energy minimization. Last, the image is eroded and dilated, and the cloning separate parts could be got effectively. For the pixels which may be partitioned wrongly, we use a tool similar to a brush to re-mark the front object or the background, and divide once again. Both subjective the evaluation criteria and the RUMA, evaluation criteria are used to evaluate the method, and the experiment results are satisfactory.
Humans
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Image Enhancement
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methods
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Image Interpretation, Computer-Assisted
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methods
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Multiple Myeloma
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classification
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Pattern Recognition, Automated
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methods
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Tumor Stem Cell Assay
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instrumentation
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methods
10.Effect of Neoadjuvant Chemotherapy Treatment on Prognosis of Patients with Advanced Gastric Cancer: a Retrospective Study.
Shu-bo TIAN ; Jian-chun YU ; Wei-ming KANG ; Zhi-qiang MA ; Xin YE ; Chao YAN ; Ya-kai HUANG
Chinese Medical Sciences Journal 2015;30(2):84-89
OBJECTIVETo evaluate the prognostic effects of neoadjuvant chemotherapy (NAC) in patients with local advanced gastric cancer.
METHODSWe retrospectively analyzed prognosis in 191 patients with advanced gastric cancer, of whom 71 were treated with NAC and 120 received surgery only between February 2007 and July 2013. Postoperative complication rate was recorded. Survival by clinicopathological features, pathological T and N stages, and histopathological tumor regression was retrospectively compared between the two groups.
RESULTSAccording to Response Evaluation Criteria in Solid Tumors, none of the 71 patients in the NAC followed by surgery group showed complete response, 36 showed partial response, 25 had stable disease, and 10 had progressive disease. The chemotherapy response rate was 50.7%; the disease control rate was 85.9%. Grade 3/4 adverse events were seen in less than 20% patients, with acceptable toxicities. No difference was found in the overall postoperative complication rates between the two groups (7 versus 22 cases, P=0.18). Median survival time was significantly different, at 54 months in the NAC combined with surgery group and 25 months in the surgery-only group (P=0.025).
CONCLUSIONIn patients with operable gastric adenocarcinomas, NAC can significantly improve overall survival without increasing surgical complications.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoadjuvant Therapy ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; drug therapy ; mortality ; pathology