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Article in Chinese | WPRIM | ID: wpr-845224


Objective: To develop an HPLC method for quantitative analysis of multi- components by single marker(QAMS)to simultaneously determine gypenoside XL,gypenoside A, gypenoside X,danshensu,salvianolic acid B,tanshinone Ⅱ A,orientin,isoorientin,vitexin and isovitexin in Compound Jinpu Tablet(CJT). Methods: HPLC was performed on a Wondasil C18 column(250 mm×4.6 mm,5 μm)at 30℃. The mobile phase was methanol-aceto-nitrile(3:1)and 0.1% formic acid aqueous solution,in a gradient elution,and the flow rate was 0.9 ml/min. The detec-tion wavelength was set at 203 nm for gypenoside XL,gypenoside A and gypenoside X,280 nm for danshensu,sal- vianolic acid B and tanshinoneⅡA,and 340 nm for orientin,isoorientin,vitexin and isovitexin. The content of the ten components in CJT was determined at first by the external standard method(ESM)using ten related standards. For the QAMS method,only tanshinone ⅡA was used as an internal standard,whereas the relative correction factors of the other nine components in CJT were determined using the internal tanshinone ⅡA as reference standard,and their content was calculated on the basis of the correction factors and the tanshinone ⅡA content determined by ESM. The QAMS method was validated by comparison of the calculated values with the measured data by ESM. Results: The 10 components all showed a good linearity within the concentration ranges of 2.848-56.96,5.304-106.08,7.776-155.52,1.248-24.96, 13.28-265.6,1.976-39.52,1.112-22.24,0.928-18.56,0.696-13.92,and 0.648-12.96 μg/ml(r≥0.9992),with the average recoveries(RSDs)of 99.35%(0.97%),99.07%(1.10%),98.81%(0.83%),97.69%(1.39%),100.05%(0.79%),97.90%(1.20%),98.21%(1.52%),97.59%(1.44%),96.93%(1.07%)and 96.99%(0.89%),respectively. No significant differences were found in the content of the nine components in CJT between the QAMS-calculated values and the ESM-measured data. Conclusion: The established QAMS method could simultaneously determine multi-compo- nents using only one standard. It is simple and accurate,which could be used for simultaneous determination of the con- tent of ten components in CJT.

Article in Chinese | WPRIM | ID: wpr-744342


Objective To understand pathogenic microorganism contamination status of medical waste sharps containers with different use time, explore the reasonable duration of service time of sharps containers, provide reference for the management of medical waste.Methods Twelve 2 L sharps containers on treatment trolleys in a tertiary first-class infectious disease specialty hospital were randomly selected, viral loads of hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) as well as bacterial colonies on inner and outer surfaces of sharps containers at different time points were detected, three unused sharps containers were taken as control at the same time.Results Sixty eluent specimens of outer surface and contents of sharps containers in trial group and control group were collected respectively at four time points (48 h, 72 h, 5 d, 7 d), no HIV and HCV were detected, and no HBV was detected in specimens of outer surface of sharps containers, HBV was detected in the eluent of contents in one sharps container 72 hours after the use, concentration of HBV was 2.20 E+01 IU/mL. Changes in bacteria in the eluent of used sharps containers: 100% of the eluent of contents in sharps containers grew bacteria on the 5 th day after use, bacterial load of the eluent of contents in sharps containers on the 7 th day after use was incalculable. Bacterial load on the outer surface of sharps containers ranged from 1 to 9 CFU/cm2. No significant changes were observed in the inner and outer surfaces of all sharps containers, and no discomfort odor emerged.Conclusion With the storage time prolonged to 7 days, bacterial colonies on the outer surface of sharps containers didn't increase significantly, HIV, HBV and HCV were not detected. It is suggested that service time of sharps containers with small production of contents should not be set compulsorily at 48 hours (even if the contents in sharps container is less than 3/4 of storage capacity after 48 hours of use).

Acta Pharmaceutica Sinica ; (12): 198-205, 2017.
Article in Chinese | WPRIM | ID: wpr-779579


It has been an active approach to screen the active ingredients in traditional Chinese medicines (TCMs) according to the affinity property between small molecule compounds and biomaterials such as cells, bacteria and proteins. On the other hand, the biomaterials can be immobilized on a solid support before the screening procedure. The immobilization method not only can maintain the biological activities of biomaterials, but also have other advantages such as high efficiency, simple operation, easy to be continuous and automatic, etc. Carrier materials (solid supports) for the immobilization including silica gel, magnetic materials, hollow fiber, and the surface plasma resonance sensor chips have been used to immobilize biomaterials and successfully applied in the screening of active ingredients from TCMs. In this paper, applications of immobilization techniques in the screening of active components from TCMs were reviewed to provide a scientific reference to the future applications.

Article in Chinese | WPRIM | ID: wpr-237214


<p><b>OBJECTIVE</b>To investigate the efficiency of multi-round fluorescence in situ hybridization (FISH) and its influencing factors in preimplantation genetic diagnosis (PGD).</p><p><b>METHODS</b>A total of 48 couples accepted PGD because of various reasons: 24 with Robertsonian translocations, 16 with reciprocal translocations, 2 with pericentric inversions, one with advanced maternal age who had a previous liveborn of Down syndrome, 3 suffered from sex chromosome abnormalities and 2 repeated spontaneous miscarriages. After 72 retrieval cycles, 432 cleavage stage embryos with more than six cells were biopsied on day three. Only intact nuclei (396) were hybridized in order to verify the chromosomal status of the individual embryos. If previous FISH has failed to give conclusive results while the nuclei remained undamaged, the nuclei were hybridized once again. A total of 870 times of hybridization were conducted to 396 nuclei. Signal identification rates of each round as well as the influence of different probes to the hybridization efficiency were compared. Factors leading to inconclusive FISH results were analyzed as well.</p><p><b>RESULTS</b>Five hundred and thirty five out of 870 hybridizations gave identifiable signals (61.5%). The second and third round FISH showed the best signals with an identification rate of 71.8% and 77.4%, respectively, which were significantly higher than those of the first round (52.8%, P < 0.01), the fourth round (55.8%, P < 0.05, P < 0.01), the fifth round (54.5%, P < 0.05) and the sixth round (27.3%, P < 0.01). The identification rate of centromere specific probe signals (CEP group) was 80.3% and the former three rounds in this group got the best quality of signals with an identification rate of 85.7%, 85.1% and 88.0%, respectively, which was significantly higher than that of the latter three rounds. The identification rate of other probe was much lower than with the CEP probe (55.2% vs. 80.3%, P < 0.01) and the best quality of signal in this group was achieved in the fifth round (72.7%), followed by the second round (66.1%) and the third round (63.8%). The identification rate of the first round (50.3%) and the sixth round (22.2%) were significantly lower compared with the second round (P < 0.01). During the 6 rounds of FISH, 335 hybridizations did not give conclusion results (38.5%, 335/870). The main cause of unidentification was weak signals (20.9%, 182/870). Other common factors included background interference (7.6%, 66/870) and failed hybridization (6.1%, 53/870). Rare causes included nucleus damage (1.8%, 16/870), nucleus loss (1.1%, 10/870) and signal split/overlap (0.9%, 8/870).</p><p><b>CONCLUSION</b>Multi-round FISH can improve the utility of single nucleus in PGD and the former three rounds have the highest efficiency. The hybridization effect of CEP is better than other probe. Poor signal quality is the common cause of unidentification results.</p>

Female , Genetic Testing , Methods , Humans , In Situ Hybridization, Fluorescence , Methods , Male , Pregnancy , Preimplantation Diagnosis , Methods , Prenatal Diagnosis , Translocation, Genetic
National Journal of Andrology ; (12): 799-804, 2011.
Article in Chinese | WPRIM | ID: wpr-305787


<p><b>OBJECTIVE</b>To investigate the role of dynactin 1 (Dctn1) in the process of mouse spermiogenesis.</p><p><b>METHODS</b>Western blot and indirect immunofluorescence were used to analyze the expression and location of Dctn1 in the mouse testis and spermatozoa. The highest efficiency of small interference RNA (siRNA) was verified by GC2-spd cell line in vitro and in vivo studies, respectively. Dctn1 siRNA mixed with the indicator (0.4% trypan blue) was injected into the seminiferous tubules of 3-week-old ICR mice through rete testis microinjection, and negative control siRNA injected into the control testes. The normal group included 3-week-old ICR mice that did not receive any treatment. Spermatozoa were collected from the cauda epididymis 3 weeks after siRNA injection for morphological analysis.</p><p><b>RESULTS</b>Dctn1 was mainly localized in the tail of spermatozoa. After interference, the sperm tail abnormality in the Dctn1 siRNA group was (23.57 +/- 0.55)%, significantly higher than (12.35 +/- 2.29)% in the control (P < 0.01, n = 3), and it was (3.37 +/- 0.69)% in the normal group.</p><p><b>CONCLUSION</b>Dctn1 plays an important role in mouse spermiogenesis, and mainly affects the formation of the tail of spermatozoa.</p>

Animals , Dynactin Complex , Male , Mice , Mice, Inbred ICR , Microinjections , Microtubule-Associated Proteins , Genetics , Metabolism , RNA, Small Interfering , Rete Testis , Metabolism , Seminiferous Tubules , Metabolism , Sperm Count , Sperm Motility , Spermatogenesis , Spermatozoa , Metabolism , Testis , Metabolism
Article in Chinese | WPRIM | ID: wpr-357599


<p><b>OBJECTIVE</b>To investigating genetic effects of workers occupationally exposed to mercury (Hg).</p><p><b>METHODS</b>The peripheral lymphocytes from 20 workers exposed to mercury and 20 controls were measured with micronucleus test, comet assay, hrpt gene mutation test and TCR gene mutation test.</p><p><b>RESULTS</b>The mean micronuclei rate(MNR) and mean micronucleated cells rate(MCR) in 20 workers were (5.90 +/- 0.91) per thousand and (5.30 +/- 0.81) per thousand, respectively while MNR and MCR in controls were (1.50 +/- 0.47) per thousand and (1.30 +/- 0.31) per thousand respectively, The difference of MNR and MCR between workers and controls was very significant (P < 0.01). The mean tail length (MTL) of workers and controls were (3.16 +/- 0.31) and (0.99 +/- 0.07) microm, respectively. The mean tail moment (MTM) of workers and controls were 1.63 +/- 0.22 and 0.39 +/- 0.03, respectively, There was a significant difference in MTL and MTM between workers and controls(P < 0.01). When the average mutation frequencies (Mfs-hprt) of hprt and (Mfs-TCR) of TCR of workers were compared with those of controls, there were not significant difference (P > 0.05).</p><p><b>CONCLUSION</b>The results of the investigation indicated that the adverse genetic effects in workers occupationally exposed to mercury could be detected.</p>

Adult , Case-Control Studies , Chemical Industry , Comet Assay , Female , Humans , Male , Mercury , Micronucleus Tests , Middle Aged , Mutation Rate , Occupational Exposure , Young Adult