1.Application of in situ amniocyte culture for prenatal diagnosis.
Daoqi HUANG ; Huayu TU ; Chaohong WANG ; Yaqun WANG ; Keting TONG ; Jiansheng ZHU
Chinese Journal of Medical Genetics 2020;37(10):1117-1119
OBJECTIVE:
To explore the value of in situ amniocyte culture for prenatal diagnosis.
METHODS:
2716 amniotic fluid samples were cultured in situ on slides. After the culture, the slides were stained, photographed and analyzed.
RESULTS:
All samples were successfully analyzed, with the success rates for primary culture and subculture being 98.42% and 1.58%, respectively. 224 samples (8.25%) were detected with chromosomal aberrations, which included 125 cases with trisomy 21, 31 with trisomy 18, 3 with trisomy 13, 4 with 45,X, 17 with 47,XXY, 5 with 47,XYY, 1 with 48,XXY,+18, 1 with 48,XXYY, 26 with structural chromosomal aberrations, and 11 with mosaicisms for aneuploidies.
CONCLUSION
In situ amniocyte culture is stable and has a high success rate, and is capable of identifying true and false mosaicisms, which can improve the accuracy of prenatal diagnosis.
2.Improvement and application of culture method for hemorrhagic amniotic fluid cells
Daoqi HUANG ; Huayu TU ; Keting TONG ; Chaohong WANG ; Jiansheng ZHU
Chinese Journal of Clinical Laboratory Science 2024;42(2):90-92
Objective To compare the success rates of different culture methods for hemorrhagic amniotic fluid.Methods Thirty-one hemorrhagic amniotic fluid samples from pregnant women who were subjected to chromosomal examination at Anhui Maternal and Child Health Hospital were collected from January 2021 to December 2022.Two culture methods,the slide in situ culture box method(re-ferred to as the slide method)and the plastic bottle culture combined with slide in situ culture box method(referred to as the combined culture method),were used for cell cultivation.All the cells were harvested and stained with Giemsa staining,then the number of eligi-ble karyotype was counted and the success rates were compared between the two methods.Results Among the 31 cases of the slide method,21 were successfully cultured with a success rate of 67.7%.For the combined culture method,all the 31 cases were success-fully cultured with a success rate of 100%.The success rate of the combined culture method was significantly higher than that of the slide method(P<0.05).Of 31 bloody amniotic fluid samples,three cases were fresh bleeding,and an average number of eligible kary-otype was 8 in the slide method and 32 in the combined culture method.Twenty-eight cases were old bleeding,and an average number of eligible karyotype was 13 in the slide method and 53 in the combined culture method.The number of eligible karyotype in the com-bined culture method was significantly higher than that of the slide method(P<0.05).Conclusion The combined culture method is suitable for the cultivation of hemorrhagic amniotic fluid samples,and should be worthy of promotion in the clinics.
3.Correlation between appearance characteristics and intrinsic quality of Gastrodiae Rhizoma.
Ping LI ; Jin-Guo XU ; De JI ; Chun-Qin MAO ; Ke-Wei ZHANG ; Yan SONG ; Tu-Lin LU
China Journal of Chinese Materia Medica 2019;44(20):4460-4466
Totally 21 batches of Gastrodiae Rhizoma pieces from various habitats with different appearance characteristics were analyzed. Five active components( gastrodin,4-hydroxybenzyl alcohol,parishin B,parishin C,and parishin A) were determined by UPLC. Polysaccharide content was determined by phenol sulfuric acid method. And the content of alcohol extracts was determined. The correlation between appearance characteristics and active components,polysaccharide content and amount of alcohol extracts of Gastrodiae Rhizoma pieces was statistically analyzed by SPSS 19. 0 software. And the five active components in the 21 Gastrodiae Rhizoma pieces were clustered by using the Ward' s method in SPSS 19. 0 software. The study found that inconspicuous keratinous Gastrodiae Rhizoma had a low content of each component,and the degree of keratin was positively correlated with the content of each component and the extract,with no significant correlation with the polysaccharide content. In the cluster analysis,the 21 batches were divided into three groups,except the second group whose gastrodin content was significantly higher than the special conditions of other groups. According to the content of the five active components,19 batches of Gastrodiae Rhizoma pieces were classified into two groups with obvious keratin and no obvious keratin. In terms of color,with the deepening of brown color,the content of each component showed a downward trend. The grading of decoction pieces of Gastrodiae Rhizoma shall give full consideration to the color and texture. And the combination of chemical composition and appearance characteristics is an indicator to evaluate the product specification standards.
Drugs, Chinese Herbal
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Gastrodia
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Rhizome
4.Effects of Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma on immune hepatic fibrosis, proliferation of HSC-T6 and expression of α-SMA and Procollagen I.
Ji ZHANG ; Yan SONG ; Qiao-Han WANG ; Lin LI ; De JI ; Wei GU ; Min HAO ; Tu-Lin LU ; Chun-Qin MAO
China Journal of Chinese Materia Medica 2017;42(13):2538-2545
To compare the effects of Curcumae Rhizoma/vinegar-processed Curcumae Rhizomaon immune hepatic fibrosis, proliferation of HSC-T6, and expressions of α-SMA and Procollagen I. The immunological liver fibrosis model was prepared through intraperitoneal injection with porcine serum 0.5 mL in each rat, twice a week, for 14 weeks. Expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN, HA and HYP, MDA in liver tissues were observed after administration of Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma (0.95, 1.90 g•kg⁻¹). The pathological changes in liver tissues were observed by HE staining. Masson staining and Sirius red staining were used to observe the expression of collagen in rat liver. HSC-T6 was cultured, and the proliferation of HSC-T6 was determined by MTT assay at different concentrations in 12, 24, 36, 48 h. The expressions of α-SMA and Procollagen I were detected by Real-time PCR. The results showed that expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN and HA in Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma groups (0.95, 1.90 g•kg⁻¹) were significantly lower than model group; in terms of effect, vinegar-processed Curcumae Rhizoma group was superior to Curcumae Rhizoma group. Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma containing serum could inhibit the proliferation of HSC-T6 in a dose-effect and time-effect manner. Expressions of α-SMA and Procollagen I in HSC-T6 were decreased after 24 h, especially in 20% vinegar-processed Curcumae Rhizoma containing serum group (P<0.01). Both Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma could reduce immune hepatic fibrosis to varying extent. Their anti-hepatic fibrosis mechanism may be correlated with inhibition of the proliferation of HSC-T6, and reduction of the formation of extracellular matrix and promotion of its degradation.
5.Quality evaluation of multi-components simultaneous determination and fingerprint of Gardeniae Fructus from different regions.
Hong-Hong CAO ; Wei-Hua YAN ; Shuang GUO ; Min HAO ; Ke-Wei ZHANG ; Tu-Lin LU ; Chun-Qin MAO ; Lin LI ; Yan SONG
China Journal of Chinese Materia Medica 2019;44(18):4026-4033
An analysis method was established by UPLC fingerprint and then applied to simultaneous determination of multiple compounds in Gardeniae Fructus from different areas in China. Samples were separated on a Waters Acquity UPLC BEH C18( 2. 1 mm×50 mm,1. 7 μm) column with 0. 1% formic acid-water and acetonitrile solution as gradient mobile phase at a flow rate of 0. 4 m L·min-1 at various wavelengths. The similarity of samples was over 0. 95 with ″Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine( 2012 edition) ″. The UPLC common fingerprints for 32 batches were established with 19 common peaks identified. The samples were divided into 3 groups analyzed by HCA and PCA. Five components were identified as the main compositions which caused the differences of chemical constituents in the samples from different areas with partial least squares discriminant analysis( PLS-DA). The content of the total components in each area was Zhejiang > Fujian > Jiangxi > Sichuan. This method was accurate and viable,could be used to evaluate the quality of Gardeniae Fructus.
China
;
Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal/standards*
;
Fruit/chemistry*
;
Gardenia/chemistry*
;
Phytochemicals/analysis*
6.Technology optimization of Gardeniae Fructus processed with ginger juice and composition changes after processing.
Hong-Hong CAO ; Wei-Hua YAN ; Shuang GUO ; Chang DENG ; Rong XUE ; Ke-Wei ZHANG ; Tu-Lin LU ; Chun-Qin MAO ; Lin LI ; De JI ; Yan SONG
China Journal of Chinese Materia Medica 2019;44(24):5413-5420
To optimize the technology of Gardeniae Fructus processed with ginger juice,establish fingerprints and simultaneously determine seven compounds( geniposidic acid,chlorogenic acid,genipin-1-β-D-gentiobioside,geniposide,rutin,crocin Ⅰ,and crocin Ⅱ) by using ultra high performance liquid chromatography( UPLC). Waters ACQUITY UPLC BEH C18( 2. 1 mm×50 mm,1. 7μm) column was used with acetonitrile and 0. 1% formic acid solution as mobile phase for gradient elution at the flow rate of 0. 4 m L·min-1. The data was comprehensively processed and analyzed with similarity evaluation,principal component analysis( PCA) and partial least squares discriminant analysis( PLS-DA) methods. Twenty common peaks were identified in this study,and the similarity of samples was over 0. 97. The results of PCA and PLS-DA showed that there were differences in chemical compositions and contents between the raw Gardeniae Fructus and those processed with ginger juice,with 9 potential differentiated chromatographic peaks. After being processed with ginger juice,the contents of chlorogenic acid,crocin Ⅰ and crocin Ⅱ were less than before and the contents of other four compositions were higher than before. The optimized preparation for Gardeniae Fructus processed with ginger juice was stable and feasible. The methods of UPLC fingerprints and simultaneous determination of seven components can be effectively carried out to distinguish Gardeniae Fructus and Gardeniae Fructus processed with ginger juice.
Carotenoids/analysis*
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Chlorogenic Acid/analysis*
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Chromatography, High Pressure Liquid
;
Drugs, Chinese Herbal/chemistry*
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Fruit/chemistry*
;
Gardenia/chemistry*
;
Zingiber officinale
;
Technology, Pharmaceutical/methods*