1.Optimization of Vacuum-belt Drying Process of Rhizoma Chuanxiong Extract
Jingxi SI ; Zhongfang PENG ; Shengbo LIU
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(05):-
Objective To explore the optimal conditions of the vacuum-belt drying process of Rhizoma Chuanxiong extract.Methods Influencing factors of the vacuum belt drying process of Rhizoma Chuanxiong extract were studied by using orthogonal test.The water content and ferulic acid content of dried product were used as the quality indicators.Results The optimal parameters of the vacuum-belt drying process were as follows:the heated region temperature being 95 ℃,the feeding speed at 10 Hz/s,and the belt speed at10 Hz/s.Conclusion The vacuum-belt drying techniquefor drying the extract brings high drying rate and high quality product.This study can provide a reasonable basis for industrial production in line with the GMP requirements.
2.Study on Refinement of Aqueous Extract of Rhizoma Chuanxiong with Inorganic Ceramic Membrane
Jingxi SI ; Zhongfang PENG ; Shengbo LIU
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To observe the microfiltration of different types of ceramic membrane for aqueous extract of Rhizoma Chuanxiong,and to select the optimal type of ceramic membranes. Methods The aqueous extract of Rhizoma Chuanxiong was microfiltered by inorganic ceramic membranes in the pore diameter of 50 nm,200 nm,and 500 nm,respectively. The characters,total solids,and active ingredients of the aqueous extract were analyzed and compared before and after microfiltration. Results The aqueous extract turned into clarified liquid from turbid liquid after microfiltration with the three types of the inorganic ceramic membrane. The removal rate for total solids was 25.7 %,26.5 %and 25.9 %,and the loss rate for ferulic acid was 12 %,6 %and 4 %,respectively for the inorganic ceramic membranes in the pore diameter of 50 nm,200 nm,and 500 nm. Conclusion Both the yield of dry solid material and ferulic acid are stable during the three middle-scale tests,indicating that he aperture ceramic membrane in the pore diameter of 200 nm is a optimal selection for the purification and refining of aqueous extract of Rhizoma Chuanxiong.
3.Effect of RNA Interference Targeting Gene Combined with Ultrasonic Irradiation and SonoVue Microbubbles on Proliferation and Apoptosis in Keratinocytes of Psoriatic Lesions.
Li-Wei RAN ; Hao WANG ; Dong LAN ; Hong-Xia JIA ; Si-Si YU
Chinese Medical Journal 2018;131(17):2097-2104
Background:
Signal transducer and activator of transcription 3 (STAT3) was strongly expressed and activated in psoriatic keratinocytes (KCs) and correlated with the severity of psoriasis. The study aimed to investigate the effects of STAT3 small interfering RNA (siRNA) combined with ultrasonic irradiation and SonoVue microbubbles on the proliferation and apoptosis in KCs of psoriatic lesions and the relative mechanisms.
Methods:
Psoriatic KCs were transfected under four experimental conditions: (1) STAT3 siRNA carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (LUS group); (2) STAT3 siRNA only carried by Lipofectamine 3000 (L group); (3) the negative control of siRNA carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (siRNA-NC); (4) not treated as Blank. Cell Counting Kit-8 assay was used to evaluate the cell proliferation. Cell cycle analysis was detected with cycle test Plus DNA reagent kit associated with flow cytometer. FITC Annexin V apoptosis detection kit associated with flow cytometer was applied for apoptosis analysis. Fluo calcium indicator associated with flow cytometer was used to analyze intracellular free calcium concentration ([Ca]). The expressions of cyclin D1 and Bcl-xL were detected both at the mRNA level by real-time reverse transcription-polymerase chain reaction (RT-PCR) and at the protein level by Western blotting. The obtained data were statistically evaluated by two-way analysis of variance.
Results:
STAT3 siRNA inhibited the growth of KCs in a time-dependent manner showing the highest proliferation inhibition in LUS group with proliferation ratio of 45.38% ± 5.85% at 72h (P < 0.05 vs. L group, siRNA-NC, or Blank). STAT3 siRNA induced an altered cell cycle distribution of KCs showing the highest increases in G2/M-phase population up to 18.06% ± 0.36% in LUS group (P < 0.05 vs. L group, siRNA-NC, or Blank). STAT3 siRNA induced late apoptosis of KCs with the highest late apoptosis percentage of 22.87% ± 1.28% in LUS group (P < 0.05 vs. L group, siRNA-NC, or Blank). STAT3 siRNA induced the elevation of [Ca]of KCs with the highest calcium fluorescence intensity mean of 1213.67 ± 60.51 in LUS group (P < 0.05 vs. L group, siRNA-NC, or Blank). STAT3 siRNA induced the downregulation of cyclin D1 and Bcl-xL expressions of KCs at mRNA and protein levels with the lowest expressions in LUS group with cyclin D1 expression of 51.81% ± 9.58% and 70.17% ± 4.22% at mRNA level and at protein level, respectively, and with Bcl-xL expression of 37.58% ± 4.92% and 64.06% ± 7.78% at mRNA level and at protein level, respectively (P < 0.05 vs. L group, siRNA-NC, or Blank).
Conclusions
STAT3 siRNA inhibited the growth and induced the apoptosis in psoriatic KCs likely partly through altering cell cycle distribution, elevating [Ca], and downregulating cyclin D1 and Bcl-xL expressions. Silencing the target gene STAT3 in psoriatic KCs with siRNA combined with ultrasonic irradiation and microbubbles would contribute to a significant innovation as a new clinical therapy for psoriasis.
Apoptosis
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Cell Line, Tumor
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Cell Proliferation
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Humans
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Keratinocytes
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Microbubbles
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Phospholipids
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Psoriasis
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therapy
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RNA Interference
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RNA, Small Interfering
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STAT3 Transcription Factor
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metabolism
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Sulfur Hexafluoride
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Ultrasonics