1.Optimization and Preliminary Evaluation the Formulation of Self-microemulsifying Drug Delivery System of Potassium Dehydroandrographolidi Succinas
Hao WANG ; Mingquan CUI ; Rongli YIN ; Zhimin CI ; Quanyi YU
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):712-718
The self-microemulsion formulation of potassium dehydroandrographolidi succinas (PDS) has been optimized and the performance in vitro has been evaluated preliminary. Kinds of prescription accessories were screened by solubility based on the emulsifying result and efficiency, particle size of emulsions. The optimal formulation composition and compatibility proportion were determined by orthogonal design and pseudo-ternary phase diagrams. The appearance, particle size, Zeta potential and stability of this formulation were also investigated. The optimized prescription of PDS was 10% MCT, 40% Tween-20 and 50% glycerol. It can spontaneously form a transparent pale blue opalescent emulsion with emulsification time 31.27 s, particle size 37.1 nm, Zata potential -17.4 mV and good stability.
2.Feasibility of spermatogonial stem cells separation with alpha 6-integrin and c-kit as specific surface makers in mice
Xianjin DU ; Ci ZHANG ; Chunxia LIU ; Linglong WANG ; Xiaoxiang YU
Chinese Journal of Tissue Engineering Research 2007;0(36):-
BACKGROUND:It is accepted that the best method for spermatogonial stem cells separation is using artificial cryptorchism model combined with surface makers.OBJECTIVE:To explore the feasibility of separation spermatogonial stem cells with ?6-integrin and c-kit as specific surface markers.DESIGN,TIME AND SETTING:The randomized control experiment was performed at the Renmin Hospital of Wuhan University from May to December 2006.MATERIALS:Forty adult,white Kunming mice with 6 weeks old were randomly divided into cryptorchidism and control groups,with 20 animals in each group.METHODS:Artificial cryptorchidism model was prepared by made an incision at the median of abdomen,and testis was pulled into abdominal cavity,which was fixed at the each side of lateral abdominal wall.There was no treatment in the control group.The single cell suspension of seminiferous epithelium was obtained by traditional two step enzyme digestion at 2-3 months after operation.FITC-conjugated anti-?6-intergrin antibody and PE-conjugated anti-c-kit antibodies were added.Then the cells with low side scatter light-scattering properties were sorted and positively stained for ?6-intergrin and negative c-kit expression.Meanwhile,the viability of the isolated cells was assessed by trypan blue staining.MAIN OUTCOME MEASURES:The morphological changes of cryptorchidism,and the sorting results of spermatogonial stem cells.RESULTS:Cell distribution in seminiferous tubule was disorder with reduced numbers.The layer and lumens were disappeared,and cell division phase could be seen in the center of tubules.Compared to the control group,the testicular cells in the cryptorchidism group were increased in the side scatterlow,Forward scatterhi areas,with figure left-upward displacement.The distribution of ?6-integrin+ and c-kit cells were deviated each other,it named that most ?6-integrin+ cell were not spermatogonial stem cells,so do the c-kit-cells.Only 2.8% of testicular cells exhibited side scatterlow,?6-integrin+,and c-kit-,which were spermatogonial stem cells in the cryptorchidism group.And trypan blue staining showed that over 95% of them were viable.CONCLUSION:Using the two surface markers to sort spermatogonial stem cells can advance the purity of the spermatogonial stem cells in cell suspension,but the specificity is insufficient.
3.The clinical effectiveness of detrusor combined triangle injection of Botulinum toxin type A plus hydrodistention in treating interstitial cystitis/bladder pain syndrome
Ci ZOU ; Dexin YU ; Yi WANG ; Li ZHAO ; Qi WANG ; Zhiqiang ZHANG ; Dongdong XIE ; Liangkuan BI ; Tao ZHANG ; Demao DING
Chinese Journal of Urology 2017;38(8):604-610
Objective To evaluate the effectiveness and security of Botulinum toxin type A (BTX-A) in treating interstitial cystitis /bladder pain syndrome,and also the effect of different site injection.Methods Sixty-nine consecutive BPS/IC patients from October 2011 to February 2016 were divided into three groups randomly,including control group (group A) and treatment group (group B and group C).There were 5 males and 64 females,with age from 23 to 66 years old (average 44.5 years old).Twentythree patients (1 male and 22 females,aged from 23 to 69 years old,with mean age of 44.6 years old) in control group (group A) underwent hydrodistention.Twenty-four patients (2 males and 22 females,aged from 27 to 65 years old,with mean age of 42.8 years old) in group B underwent BTX-A detrusor combined triangle injection plus hydrodistention.Twenty-two patients (2 males and 20 females,aged from 30 to 68 years old,with mean age of 44.3 years old) in group C underwent BTX-A triangle injection alone plus hydrodistention.The parameters such as daytime frequency,nocturia,visual analogue scale/score (VAS),functional cystemetric volume (FCV),post-void residual volume (PVR),QOL score,maximal bladder capacity (MBC),interstitial cystitis symtom index (ICSI),interstitial cystitisproblem index (ICPI),Qmax,and Hamilton anxiey scale (HAMA) score between pre-treatment and 1,3,6 and 9 months after therapy in the three groups were compared.Results There were no serious complications observed in the three groups.All parameters 1 month after therapy were obviously superior to that of pretreatment.The efficacy in control group decreased significantly 3 months after treatment except nocturia (3.0 vs.5.0),daytime frequency(7.0 vs.14.0)and Q (14.0 ml/s vs.13.0 ml/s).However,all parameters in research groups except PVR were still obviously superior to pretherapy.The parameters except Q and nocturia in group B had statistically significant difference from contemporaneous group A (all P < 0.05).However,the parameters except Q nocturia and somatic anxiety score in group C had statistically significant difference from contemporaneous group A (all P < 0.05).VAS (2.0 vs.3.0) and somatic anxiety score (6.0 vs.10.0) in group B were superior to group C (all P < 0.05).When it came to 6 months after therapy,urinary frequence and urgency and pain symptoms were the same to pretherapy and all parameters had no statistically significant difference between pretherapy and after therapy in group A(all P > 0.05).But all parameters except PVR in research group was still superior to pretherapy.the parameters except PVR,Q and QOL in group B had statistically significant difference from contemporaneous group A (all P < 0.05).However,The parameters except PVR,Q MBC,ICSI and QOL in group C had statistically significant difference from contemporaneous group A (all P < 0.05).Meanwhile,efficacy in group B was better than group C in term of ICPI (6.0 vs.8.0) and somatic anxiety score (7.0 vs.10.0) (all P < 0.05).The efficacy decreased significantly 9 months after treatment in both group B and C,with no statistically significant difference compared with that of pretreatment.ICSI(10.0 vs.13.0),ICPI(9.0 vs.13.0),QOL(5.0 vs.6.0)in group B,and QOL(5.0 vs.6.0)in group C had statistically significant difference compared with the contemporary parameters in group A.ICSI(10.0 vs.12.0),MBC(285.0 ml vs.237.5 ml) in group B was better than that in group C (P < 0.05).Conclusions Symptoms in IC/BPS patients can be alleviated significantly by detrusor BTX-A injection plus hydrodistention.Quality of life can be improved remarkably and HAMA scores can be reduced significantly after treatment.Thus,it's an effective therapeutic mnethod for IC/BPS,and detrusor combined triangle injection can provide a better effect than single triangle injection.
4.Construction of fetal mesenchymal stem cell cDNA subtractive library.
Li YANG ; Dong-Mei WANG ; Liang LI ; Ci-Xian BAI ; Hua CAO ; Ting-Yu LI ; Xue-Tao PEI
Journal of Experimental Hematology 2002;10(2):89-92
UNLABELLEDTo identify differentially expressed genes between fetal mesenchymal stem cell (MSC) and adult MSC, especially specified genes expressed in fetal MSC, a cDNA subtractive library of fetal MSC was constructed using suppression subtractive hybridization (SSH) technique. At first, total RNA was isolated from fetal and adult MSC. Using SMART PCR synthesis method, single-strand and double-strand cDNAs were synthesized. After Rsa I digestion, fetal MSC cDNAs were divided into two groups and ligated to adaptor 1 and adaptor 2 respectively. Results showed that the amplified library contains 890 clones. Analysis of 890 clones with PCR demonstrated that 768 clones were positive. The positive rate is 86.3%. The size of inserted fragments in these positive clones was between 0.2 - 1 kb, with an average of 400 - 600 bp.
CONCLUSIONSSH is a convenient and effective method for screening differentially expressed genes. The constructed cDNA subtractive library of fetal MSC cDNA lays solid foundation for screening and cloning new and specific function related genes of fetal MSC.
Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Fetus ; Gene Library ; Humans ; Mesoderm ; cytology ; metabolism ; Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism
5.Sorting of spermatogonial stem cells in mice.
Chun-xia LIU ; Ci ZHANG ; Ling-long WANG ; Xiao-xiang YU
National Journal of Andrology 2007;13(1):24-26
OBJECTIVETo search for an effective method for enriching spermatogonial stem cells in mice.
METHODSBilateral artificial cryptorchidism was performed on 20 six-week old male Kunming mice. Three months after the operation, the testes were removed and single cell suspension prepared by two-step enzyme digestion. FITC-conjugated anti-alpha6-integrin antibody and PE-conjugated anti-c-kit antibody were added for adequate time on ice. Then the cells with low side scatter light-scattering properties were sorted and positively stained for alpha6-integrin and negative c-kit expression. And the viability of the isolated cells was assessed by trypan blue exclusion.
RESULTSThe sorted spermatogonial stem cells constitute 2.8% of the testis cells and over 95% of them were viable.
CONCLUSIONFACS can be used to isolate quantities of viable spermatogonial stem cells.
Animals ; Cell Separation ; methods ; Cryptorchidism ; Disease Models, Animal ; Flow Cytometry ; Fluorescence ; Male ; Mice ; Mice, Inbred Strains ; Random Allocation ; Spermatogonia ; cytology ; Stem Cells ; cytology
6.Effect of a simulated He-O2 saturation dive to 65 msw on oxidative stress in humans.
Ci LI ; Jia HE ; Min-Hang LIU ; Yu-Ming LIU ; Heng-Rong YUAN ; Xiao-Bo LIU ; Yan WANG ; Tao YANG ; Yi-Qun FANG ; Wei-Bing XIAO
Chinese Journal of Applied Physiology 2011;27(2):250-252
OBJECTIVETo investigate whether a simulated He-O2 saturation dive to 65 msw would affect oxidative balance in humans.
METHODSSeven divers participated in a simulated saturation dive to 0.75 MPa (65 msw). 24-h urine samples were collected twice before, twice during, and twice after the dive, then were analyzed for contents of superoxide dismutase (SOD), malondialdehyde (MDA), total amino acid (T-AA) and total anti-oxidant capacity (T-AOC). Meanwhile, total urine volume and body weight were measured.
RESULTSThe content of T-AA was higher. (P < 0.05) than the base value in final decompression, but reverse to normal at one week after decompression. There were no changes in contents of SOD, MDA and T-AOC during and after the dive compared with their basic value. Total urine volume was lower (P < 0.05, vs basic value) at first day in chamber, then returned to normal. Body weight gradually increased after compression till the end of decompression (higher than basic value, P < 0.05).
CONCLUSIONThese data indicate that simulated saturation dive to 65 msw may not induce obvious oxidative damage, but it is necessary to monitor 24-h urine volume and oxidative sress by time in order to prevent from tissue injury.
Adult ; Amino Acids ; urine ; Decompression ; Diving ; physiology ; Helium ; chemistry ; Humans ; Male ; Malondialdehyde ; urine ; Oxidative Stress ; physiology ; Oxygen ; adverse effects ; chemistry
7.The down-regulation of miR-129 in breast cancer and its effect on breast cancer migration and motility.
Qiu-Yu WANG ; Jun TANG ; Ci-Xiang ZHOU ; Qian ZHAO
Acta Physiologica Sinica 2012;64(4):403-411
To search the microRNAs (miRNA) which suppress metastasis of breast cancer, we utilize three well known micoRNA target prediction programs, Targetscan, Pictar and miRanda, to select the microRNAs that target the genes related to tumor metastasis. We chose MDA-MB-231 with high metastasis ability as the model to evaluate the effect of miRNAs on cell motility through Transwell migration assay. After the first round of screening, miR-129 is found to significantly inhibit the migration of MDA-MB-231 both in Transwell migration assay and wound healing assay. Furthermore, miR-129 also shows great suppressive ability to cell mobility and migration in another two breast cancer cell lines BT549 and MDA-MB-435s. Most importantly, miR-129 is down-regulated both in breast cancer tissues compared with the paired adjacent normal breast tissues, and in breast cancer cell lines compared with normal breast epithelial cell MCF10A (P < 0.05). These results indicate that over-expression of miR-129 could inhibit breast cancer motility and migration, and the down-regulation of miR-129 may participate in the breast cancer migration and metastasis.
Breast Neoplasms
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pathology
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Cell Line, Tumor
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Cell Movement
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Down-Regulation
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Humans
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MicroRNAs
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metabolism
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Neoplasm Invasiveness
8.Analysis of 8-hydroxy-2'-deoxyguanosine in human urine by ultra-high performance liquid chromatography mass spectrometer method
Zhong LIU ; Zhigang YU ; Yanhua WANG ; Ying CI ; Zhan SUN ; Haitao JIAO ; Huawei DUAN
Chinese Journal of Preventive Medicine 2016;50(4):357-361
Objective To develop an ultra?high performance liquid chromatography mass spectrometer method for the rapid determination of 8?hydroxy?2'?deoxyguanosine (8?OHdG) in human urines. Methods 8?OHdG standard solution with the concentration from 0.01 to 0.1μg/ml was formulated. The solution was implanted into ion source with a rate of 7μl/min, the mass?to?charge ratio of parent ion and product ions, and ion mass to charge ratio was identified. The mass spectrum parameters of each Ion pairs, such as DP, EP and EXP, were gradually optimized. The urine sample with a concentration of 10.0μg/L was detected, and the pH of the sample was adjusted using 1 mol/L ammonium formate and formic acid solution with a volume ratio of 5∶1, 4∶1, 3∶1, 2∶1, and 1∶1. It was tested using three different polarity of SPE, i.e.:HLB, MCX, and MAX. The elution effect of methanol and water mixture with the proportion of 90∶10, 80∶20, 50:50, 20:80, and 10:90 were tested, and then acetonitrile and water mixture with the proportion of 90∶10, 80∶20, 50∶50, 20∶80, 10∶90 were also tested. The standard curve was constructed using the ratio of a standard series application fluid concentration to corresponding compounds quantitative ion liquid concentration of the peak area. The detection limit was determined as 3 times of the signal to noise ratio corresponding to the concentration of 8?OHdG, and the quantitative lower limit was determined as 10 times of the signal to noise ratio corresponding to the concentration of 8?OHdG. The blank urine spiked recovery method was used to evaluate the precision and recovery rate. Results The mass to charge ratio of parent ion was 284.1 and the product ions was 168.1, 140.1, 123.0, and 112.0, respectively. The collision voltage of quantitative ion?pair 284.1/168.1 was 18 V, the 284.1/140.1 collision voltage was 42 V, the 284.1/123.0 collision voltage was 48 V, and the 284.1/112.0 collision voltage was 53 V. The recovery rate was the highest (87.9%-104.3%) when the pH of urine was adjusted by a 10 ml 1 mol/L ammonium formate solution, 2 ml of formic acid, 88 ml of water are mixed with the sample solution volume ratio of 1∶5, and then purified with 3 ml of methanol and 3 ml water activated HLB extraction column. Within 1.0-100.0 μg/L concentration range, 8?OHdG standard application solution test results showed a good linear relationship. The regression equation was y=1.25x+0.74, and the correlation coefficient was r=0.999 5. The detection limit was 0.2μg/L, and the limit of quantification was 0.7μg/L. The method of recovery rate was in the range of 87.9%to 104.3%, the precision was in the range from 1.5% to 3.7% and inter?assay precision was in the range from 1.6% to 5.4%. Conclusion The method developed in this study had high sensitivity, good precision and accuracy, and a wide range of testing concentrations.
9.Analysis of 8-hydroxy-2'-deoxyguanosine in human urine by ultra-high performance liquid chromatography mass spectrometer method
Zhong LIU ; Zhigang YU ; Yanhua WANG ; Ying CI ; Zhan SUN ; Haitao JIAO ; Huawei DUAN
Chinese Journal of Preventive Medicine 2016;50(4):357-361
Objective To develop an ultra?high performance liquid chromatography mass spectrometer method for the rapid determination of 8?hydroxy?2'?deoxyguanosine (8?OHdG) in human urines. Methods 8?OHdG standard solution with the concentration from 0.01 to 0.1μg/ml was formulated. The solution was implanted into ion source with a rate of 7μl/min, the mass?to?charge ratio of parent ion and product ions, and ion mass to charge ratio was identified. The mass spectrum parameters of each Ion pairs, such as DP, EP and EXP, were gradually optimized. The urine sample with a concentration of 10.0μg/L was detected, and the pH of the sample was adjusted using 1 mol/L ammonium formate and formic acid solution with a volume ratio of 5∶1, 4∶1, 3∶1, 2∶1, and 1∶1. It was tested using three different polarity of SPE, i.e.:HLB, MCX, and MAX. The elution effect of methanol and water mixture with the proportion of 90∶10, 80∶20, 50:50, 20:80, and 10:90 were tested, and then acetonitrile and water mixture with the proportion of 90∶10, 80∶20, 50∶50, 20∶80, 10∶90 were also tested. The standard curve was constructed using the ratio of a standard series application fluid concentration to corresponding compounds quantitative ion liquid concentration of the peak area. The detection limit was determined as 3 times of the signal to noise ratio corresponding to the concentration of 8?OHdG, and the quantitative lower limit was determined as 10 times of the signal to noise ratio corresponding to the concentration of 8?OHdG. The blank urine spiked recovery method was used to evaluate the precision and recovery rate. Results The mass to charge ratio of parent ion was 284.1 and the product ions was 168.1, 140.1, 123.0, and 112.0, respectively. The collision voltage of quantitative ion?pair 284.1/168.1 was 18 V, the 284.1/140.1 collision voltage was 42 V, the 284.1/123.0 collision voltage was 48 V, and the 284.1/112.0 collision voltage was 53 V. The recovery rate was the highest (87.9%-104.3%) when the pH of urine was adjusted by a 10 ml 1 mol/L ammonium formate solution, 2 ml of formic acid, 88 ml of water are mixed with the sample solution volume ratio of 1∶5, and then purified with 3 ml of methanol and 3 ml water activated HLB extraction column. Within 1.0-100.0 μg/L concentration range, 8?OHdG standard application solution test results showed a good linear relationship. The regression equation was y=1.25x+0.74, and the correlation coefficient was r=0.999 5. The detection limit was 0.2μg/L, and the limit of quantification was 0.7μg/L. The method of recovery rate was in the range of 87.9%to 104.3%, the precision was in the range from 1.5% to 3.7% and inter?assay precision was in the range from 1.6% to 5.4%. Conclusion The method developed in this study had high sensitivity, good precision and accuracy, and a wide range of testing concentrations.
10.Nearly and long -term clinical effect of exenatide in obese adolescents with abnormal glucose metabolism
Zhao-Sui HUANG ; Zhao-Xuan HUANG ; YANG-jiao ; Li-Li LUAN ; Ci-Yu WANG ; Hong-Ying YANG
The Chinese Journal of Clinical Pharmacology 2015;(14):1390-1392
Objective To investigate the nearly and long-term effect of exenatide in obese adolescents with abnormal glucose metabolism. Methods A total of 92 cases of obese adolescents with abnormal glucose metabolism were randomly divided into two groups.Patients in the control group (45 cases) were treated with metformin 0.5 g tid.And those in the trial group (47 cases) were treated with metformin 0.5 g tid com-bined with exenatide ih for 6 months.The data of body weight, waist circumference, body mass index ( BMI) , fasting plasma glucose ( FPG) , 2-hour postprandial blood glucose ( 2 h PG ) , glycated hemoglobin ( HbA1 c ) , systolic blood pressure ( SBP ) , diastolic blood pressure ( DBP) , triglyceride ( TG) , total cholesterol ( TC) , high density lipopro-tein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), and fasting insulin (FINS), insulin resistance (HOMA-IR), as well as pancreatic β-cell function index ( HOMA-β) were determined before and after the treatment between two groups. The incidence rate of diabetes and hypertension were observed after three years in long-term study.Results There were significant decreases in the body weight,waist circumference and BMI in trial group ( P<0.01 ) .Compared with those in control group and themselves before treatment, there were significant decreases in 2h PG、HbA1c ( P <0.05 ) .The data of TC, LDL -C, TG were decreased in trial group (P<0.05).The SBP was significantly decreased in trial group, compared with those in con-trol group, there was statistical significant difference (P<0.05).The FINS, ISI, HOMA-IR were decreased in trial group while the HOMA-βwas increased ( P<0.01 ) , compared with those in control group and themselves before treatment, the difference was statistically significant ( P <0.05 ) .Conclusion Exenatide can significantly reduce body weight, improve endocrine metabolism and reduce the incidence of metabolic abnormalities disease caused by obesity.