Neurophysiological and neuroimaging markers of repetitive transcranial magnetic stimulation treatment response in major depressive disorder: A systematic review and meta-analysis of predictive modeling studies.

Predicting repetitive transcranial magnetic stimulation (rTMS) treatment outcomes in major depressive disorder (MDD) could reduce the financial and psychological risks of treatment failure. We systematically reviewed and meta-analyzed studies that leveraged neurophysiological and neuroimaging markers to predict rTMS response in MDD. Five databases were searched from inception to May 25, 2023. The primary meta-analytic outcome was predictive accuracy pooled from classification models. Regression models were summarized qualitatively. A promising marker was identified if it showed a sensitivity and specificity of 80% or higher in at least two independent studies. Searching yielded 36 studies. Twenty-two classification modeling studies produced an estimated area under the summary receiver operating characteristic curve of 0.87 (95% CI = 0.83-0.92), with 86.8% sensitivity (95% CI = 80.6-91.2%) and 81.9% specificity (95% CI = 76.1-86.4%). Frontal theta cordance measured by electroencephalography is closest to proof of concept. Predicting rTMS response using neurophysiological and neuroimaging markers is promising for clinical decision-making. However, replications by different research groups are needed to establish rigorous markers.

Mechanism study of platelet derived growth factor receptor alpha on the bidirectional differentiation regulation of glioma-associated oncogene homolog 1-positive mesenchymal stem cells in mice / 中华口腔医学杂志

Objective: To investigate the role of platelet derived growth factor receptor alpha (PDGFRα) on bidirectional differentiation of glioma-associated oncogene homolog 1-positive mesenchymal stem cells (Gli1+-MSC). Methods: Breeding double reporter transgenic mice ROSAmT/mG/Gli1-CreERt2/PDGFRαfl (Experimental group) and ROSAmT/mG/Gli1-CreERt2 (Control group), 20 mice in each of the two groups at four weeks of age were selected, MSC were isolated from the mouse aortic epithelium. After tamoxifen inducement, the two groups of Gli1+-MSC were screened by green fluorescent protein (GFP) labeling and flow cytometry sorting. PDGFRα was conditionally knocked out in the experimental group, and the control group Gli1+-MSC expressed PDGFRα normally. The two groups of Gli1+-MSC were subjected to adipogenic induction and fibrogenic induction, the Western blotting was performed to detect PDGFRα, adipocyte markers [perilipin and CCAAT/enhancer binding protein alpha (C/EBPα)] and fibrogenic markers [alpha smooth muscle actin (α-SMA) and fibroblast-specific protein 1 (FSP-1)] and semi-quantitative analysis was performed. The degree of cellular adipose differentiation after bidirectional induction of Gli1+-MSC in both groups was observed by oil red O staining and analyzed semi-quantitatively. Results: After tamoxifen induction, Gli1+-MSC could be accurately isolated from flow cytometry by GFP labeling. Via adipogenic differentiation, the expression of PDGFRα in the experimental group (0.017±0.002) was significantly lower than that in the control group (0.184±0.012) (t=25.48，P=0.002). The protein expressions of perilipin (3.138±0.414) and C/EBPα (3.565±0.289) were significantly higher than those in the control group (2.312±0.218 and 2.179±0.103, respectively) (t=6.21，P=0.025；t=6.69，P=0.022). Thus, the knock-out of PDGFRα enhanced the adipogenic differentiation ability of Gli1+-MSC. After fibrogenesis induction, the protein expressions of PDGFRα, α-SMA and FSP-1 in the experimental group (0.030±0.001, 0.932±0.177 and 0.276±0.020, respectively) were significantly lower than those in the control group (0.439±0.006, 1.352±0.170 and 0.835±0.097, respectively) (t=149.40, P<0.001; t=66.38,P<0.001; t=11.41,P<0.08). This suggested that the knock-out of PDGFRα significantly inhibited Gli1+-MSC differentiation toward fibroblasts. After bidirectional induction, significantly less adipocyte formation was seen in the control group and more in the experimental group. Quantitative analysis showed that the amount of oil red O staining in the experimental group (0.461±0.042) was significantly higher than that in the control group (0.017±0.007) after bidirectional induction (t=23.20, P<0.01). Conclusions: PDGFRα plays an important role in the regulation of bidirectional differentiation of vascular adventitial Gli1+-MSC.

Effective Aging Inhibition of the Thermoplastic Corn Starch Films through the Use of Green Hybrid Filler.

Recently, hybrid fillers have been widely used to improve the properties of biopolymers. The synergistic effects of the hybrid fillers can have a positive impact on biopolymers, including thermoplastic corn starch film (TPCS). In this communication, we highlight the effectiveness of hybrid fillers in inhibiting the aging process of TPCS. The TPCS, thermoplastic corn starch composite films (TPCS-C), and hybrid thermoplastic corn starch composite film (TPCS-HC) were stored for 3 months to study the effect of hybrid filler on the starch retrogradation. TPCS-C and TPCS-HC were prepared by casting method with 5 wt% of fillers: nanocellulose (NC) and bentonite (BT). The alteration of the mechanical properties, aging behavior, and crystalline structure of the films were analyzed through the tensile test, Fourier transform infrared (FTIR), X-ray diffraction (XRD), differential scanning calorimetry (DSC), and water absorption analysis. The obtained data were correlated to each other to analyze the retrogradation of the TPCS, which is the main factor that contributes to the aging process of the biopolymer. Results signify that incorporating the hybrid filler (NC + BT) in the TPCS/4BT1NC films has effectively prevented retrogradation of the starch molecules after being stored for 3 months. On the contrary, the virgin TPCS film showed the highest degree of retrogradation resulting in a significant decrement in the film's flexibility. These findings proved the capability of the green hybrid filler in inhibiting the aging of the TPCS.

Circ-CCNB1 Modulates Trophoblast Proliferation and Invasion in Spontaneous Abortion by Regulating miR-223/SIAH1 axis.

CONTEXT: Spontaneous abortion (SA) is a common disorder in early pregnancy. Circular RNAs (circRNAs) have been reported to exert important regulatory effects on trophoblast function and embryo development. OBJECTIVE: The aim of this study was to explore whether and how circRNAs regulate trophoblast function in SA during early pregnancy. METHODS: Cell proliferation, 5-bromo-2-deoxyuridine (BrdU) staining, Transwell, immunofluorescence, Western blot, RNA pull-down, and dual luciferase reporter assays were performed to investigate the effect of circRNA cyclin B1 (circ-CCNB1) on trophoblast function in HTR-8/SVneo and JEG-3 cells. RESULTS: An in vitro study demonstrated that upregulation of circ-CCNB1 significantly inhibited trophoblast proliferation and invasion compared with the controls using HTR-8/SVneo and JEG-3 cells, respectively. Moreover, miR-223 was downregulated in the villous tissues of patients with SA and was further predicted and shown to negatively interact with circ-CCNB1, which is involved in trophoblast proliferation and invasion. Using bioinformatics tools and subsequent RNA pull-down and dual luciferase assays, we found that miR-223 directly targets seven in absentia homolog-1 (SIAH1) and that upregulation of miR-223 decreased circ-CCNB1-induced SIAH1 expression levels in HTR-8/SVneo cells. Interestingly, upregulation of circ-CCNB1 suppressed trophoblast proliferation and invasion through inhibition of CCNB1 nuclear translocation induced by SIAH1. Downregulation of SIAH1 enhanced circ-CCNB1-suppressed CCNB1 nuclear protein expression in trophoblast cells. CONCLUSION: Circ-CCNB1 served as a modulator of trophoblast proliferation and invasion by sponging miR-223, thus forming a regulatory network of circ-CCNB1/miR-223/SIAH1 in modulating CCNB1 nuclear translocation, which enabled us to elucidate the molecular mechanisms involved in normal embryo implantation or in SA.

Investigation on the Awareness and Behavior of Primary School Students on Sunscreen Use in Beijing.

Objective: This study aims to understand primary school students' behavior and awareness of sun exposure and sunscreen in Beijing, China. Methods: A questionnaire survey was conducted of 232 students in grades 4-6 at a public primary school in Beijing, including 129 boys (55.6%) and 103 girls (44.4%). The contents of the questionnaire included awareness about ultraviolet rays, sun exposure, sunscreen habits, and the understanding and use of sunscreen. Results: The majority of the subjects (75.0%) said they had never been sunburned, and 26.3% had never been tanned. Only 7.3% of the primary school students had a comprehensive and correct understanding that sunlight will burn, cause cancer, tan, and age the skin. Sunscreen (47.8%), sunshades (47.4%), sun hats (44.4%), avoiding going out at noon (37.5%), and sunglasses (30.2%) were the most frequently used sunscreen means. Furthermore, 47.8% of primary school students used topical sunscreen for light protection, higher than other measures. The proportion of girls using sunscreen at least once a day was more than boys. Conclusion: Primary school students in Beijing, China, do not have a comprehensive understanding of ultraviolet rays, and there are deficiencies in protective behavior. Their awareness and use of sunscreen need to be improved.

Highly-fluorescent carbon dots grown onto dendritic silica nanospheres for anthrax protective antigen detection.

Direct synthesis of carbon dots on uniform mesoporous nanospheres is an ideal way to impart fluorescence properties to the nanomaterials and retain its original uniformity. Carbon dot-based nanospheres with high quantum yield (aqueous solution, 89.3%) were synthesized by the one-step hydrothermal treatment of sodium citrate and dendritic silica spheres grafted with N-ß-(aminoethyl)-γ-aminopropyltrimethoxysilane. Its excellent chemical properties such as fluorescence, stability, homogeneity and dispersion enable it to achieve a sensitive, specific, rapid and low-cost detection of anthrax protective antigen when used as a signal for immunochromatography.

CdSe/ZnS quantum dot-encoded maleic anhydride-grafted PLA microspheres prepared through membrane emulsification for multiplexed immunoassays of tumor markers.

Early diagnosis of tumor markers is of great importance for the successful treatment of cancer. As a high-throughput and high-sensitivity detection technology, liquid suspension biochips based on quantum dot (QD) encoded microspheres have been widely used in the immunodetection of tumor markers. In this work, maleic anhydride grafted PLA (PLA-MA) microspheres based on quantum dot encoding were used as carriers for liquid phase suspension biochips for the immunoassay of tumor markers. PLA-MA fluorescent beads are prepared by embedding CdSe/ZnS quantum dots in PLA-MA using Shirasu porous glass (SPG) membrane emulsification technology, which has high fluorescence intensity, good stability, and good dispersion. Fluorescent immunoassays on dipsticks found that PLA-MA microspheres have high biological activity and good stability, which is conducive to immunoassays. Based on this, using the characteristics of CdSe/ZnS quantum dots and flow cytometry, monochromatic and two-color coding methods were developed, and 9 distinguishable coding beads were prepared. The results showed that PLA-MA fluorescent microspheres exhibited good biocompatibility, stable coding signals, low background noise, and low detection limits when performing quaternary immunoassays on tumor markers CA125, CA199, CA724, and CEA by CdSe/ZnS QD-encoded PLA-MA microsphere binding flow cytometry.

Immunoassay of SARS-CoV-2 nucleocapsid proteins using novel red emission-enhanced carbon dot-based silica spheres.

It is imminent to develop a new type of rapid COVID-19 detection method with high sensitivity. Here, we used novel red emission-enhanced carbon dot (CD)-based silica (RCS) spheres as the signals of lateral flow immunochromatography (LFI) to ultrasensitively detect novel severe acute respiratory syndrome coronavirus 2 nucleocapsid proteins (SARS-CoV-2 NPs). The red emission of CDs can be enhanced and enriched in silica spheres by a simple way. The amino ends of the N-ß-(aminoethyl)-γ-aminopropyltrimethoxy anchor carboxyl-rich CDs and enhance the red emission, while the other end is embedded in the silica carrier. Then, the composite silica spheres werecoated with 3-(triethylsilyl) propylamine to protect the CDs, promote bioconjugation and obtain RCS spheres. The optimal emission peaks of the aqueous solution and the solid state of RCS spheres were at 634 nm and 638 nm, respectively, with quantum yields (QYs) of 48.5% and 35.7%, respectively. Their red emission has a wide excitation range (from the ultraviolet region to the red region), and the best excitation wavelength is about 580 nm. Two fluorescence detection modes of the RCS-LFI technology for the SARS-CoV-2 NP assay are available: the simple mode of observation under ultraviolet light has a sensitivity of 100 pg mL-1; the advanced mode of detection under a fluorescence microscope has a sensitivity of 10 pg mL-1. This assay also exhibits the advantages of fast detection speed, high specificity, and simple operation. In addition, the feasibility of this method in actual sample detection was verified in human serum by the standard-addition method, and the results show that the method has excellent practicability. We believe that this method will be a valuable supplement for the diagnosis of COVID-19.

Genome-wide identification and phylogenetic relationships of the Hsp70 gene family of Aegilops tauschii, wild emmer wheat (Triticum dicoccoides) and bread wheat (Triticum aestivum).

Heat shock protein 70 (Hsp70) plays an important role in plant development. It is closely related to the physiological process of cell development and the response to abiotic and biological stress. However, the classification and evolution of Hsp70 genes in bread wheat, wild emmer wheat and Aegilops tauschii are still unclear. Therefore, this study conducted a comprehensive bioinformatics analysis of Hsp70 gene in three species. Among these three species, 113, 79 and 36 Hsp70 genes were identified. They are divided into six subfamilies. Group vi-1 is different from Arabidopsis thaliana. It may be the result of early evolutionary segregation. The number of exons in different subfamilies (from 1 to 13) was different, but the distribution patterns of exons / introns in the same subfamily were similar. The results of Hsp70 promoter region analysis showed that the cis-regulatory elements of A. tauschii and wild emmer wheat were different from those of wheat. In addition, CpG island proportion of wild emmer Hsp70 was higher than that of wheat, which may be the molecular basis of heat resistance of wild wheat relative to cultivated wheat. Further comprehensive analysis of chromosome location and repeat events of Hsp70 gene showed that whole-genome duplication and tandem duplication events contributed to the evolution and expansion of Hsp70 gene in wheat. The results of non-synonymous substitution and synonymous substitution analysis showed that Hsp70 genes of three species had undergone purification selection. The expression profile analysis showed that Hsp70 gene was highly expressed in the roots during the vegetative growth period. In addition, TaHsp70 gene was highly expressed under various stress. The identification, classification and evolution of Hsp70 in wheat and its relatives provided a basis for further research on its evolution and its molecular mechanism in response to stress. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02639-5.

On the Use of OPEFB-Derived Microcrystalline Cellulose and Nano-Bentonite for Development of Thermoplastic Starch Hybrid Bio-Composites with Improved Performance.

Thermoplastic starch (TPS) hybrid bio-composite films containing microcrystalline cellulose (C) and nano-bentonite (B) as hybrid fillers were studied to replace the conventional non-degradable plastic in packaging applications. Raw oil palm empty fruit bunch (OPEFB) was subjected to chemical treatment and acid hydrolysis to obtain C filler. B filler was ultra-sonicated for better dispersion in the TPS films to improve the filler-matrix interactions. The morphology and structure of fillers were characterized by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). TPS hybrid bio-composite films were produced by the casting method with different ratios of B and C fillers. The best ratio of B/C was determined through the data of the tensile test. FTIR analysis proved the molecular interactions between the TPS and the hybrid fillers due to the presence of polar groups in their structure. XRD analysis confirmed the intercalation of the TPS chains between the B inter-platelets as a result of well-developed interactions between the TPS and hybrid fillers. SEM images suggested that more plastic deformation occurred in the fractured surface of the TPS hybrid bio-composite film due to the higher degree of stretching after being subjected to tensile loading. Overall, the results indicate that incorporating the hybrid B/C fillers could tremendously improve the mechanical properties of the films. The best ratio of B/C in the TPS was found to be 4:1, in which the tensile strength (8.52MPa), Young's modulus (42.0 MPa), elongation at break (116.4%) and tensile toughness of the film were increased by 92%, 146%, 156% and 338%, respectively. The significantly improved strength, modulus, flexibility and toughness of the film indicate the benefits of using the hybrid fillers, since these features are useful for the development of sustainable flexible packaging film.

Luminous silica colloids with carbon dot incorporation for sensitive immunochromatographic assay of Zika virus.

Here a novel strategy is reported of assembling silanized carbon dots (CDs) with porous silica templates to form fluorescent CD-based silica (FCS) colloids with uniformly packed CDs throughout the silica matrix. Dendritic silica spheres with highly accessible central-radial pores are adopted as a powerful absorbent host, which can form Si-O bonds with silane to directly fix the silanized CDs. The appropriate loading content of CDs on the inner surface of dendritic silica spheres is beneficial for the maximum fluorescence intensity of FCS colloids. High-quality silanized CDs endow multiple CD embedded silica spheres with excellent properties, including good fluorescence performance, excellent colloidal/optical stabilities and convenient biofunctionalization. The integration of these FCS colloids with a lateral flow strip platform provides an ultra-sensitive, specific and robust immunoassay method for the Zika NS1 protein with a visual detection limit of 10 pg mL-1, and has been successfully applied to the detection of Zika virus in clinical samples. In addition, we also prepared conventional Au NP-based lateral flow test strips and applied them to the detection of Zika NS1 protein. By comparison, the detection limit of immunofluorescent CD-based silica (iFCS)-based lateral flow test strips is 100-fold lower than that of Au NP-based lateral flow strips.

Ultrasensitive Detection of Severe Fever with Thrombocytopenia Syndrome Virus Based on Immunofluorescent Carbon Dots/SiO2 Nanosphere-Based Lateral Flow Assay.

Sensitive detection of severe fever with thrombocytopenia syndrome virus (SFTSV) by a point-of-care assay is of great significance for promoting clinical diagnosis. In this work, ultrasensitive detection of SFTSV was achieved by using fluorescent carbon dots/SiO2 nanospheres (CSNs) as reporters for a lateral flow assay. The prepared CSNs were resistant to extreme environments and had strong stability. The uniform CSNs with the size of about 200 nm were obtained by differential centrifugation. Their absolute quantum yields in the aqueous and solid phases are 56.3 and 36.6%, respectively. The excellent fluorescent properties of CSNs make the test strips more sensitive and have a longer assay lifetime. Thus, the visual detection limit of the lateral flow test strip based on immunofluorescent CSN (iCSN) was as low as 10 pg/mL SFTSV nucleoprotein. The sensitivity of this assay is 2 orders of magnitude higher than that of the colloidal gold-based lateral flow test strip. Besides, the assay owns good reproducibility and high specificity. Then, iCSN-based lateral flow test strips were evaluated in real samples of human serum of patients with satisfactory results. Furthermore, this assay has a general prospect for other fluorescent immunochromatography applications.

Treponema pallidum-specific antibody expression for the diagnosis of different stages of syphilis.

BACKGROUND: Tp15, Tp17, Tp45, and Tp47 are outer-membrane proteins found in Treponema pallidum, the etiologic agent of syphilis. These proteins are potent antigens and are potential markers for the serological detection of syphilis. The present study analyzed antibodies to these protein antigens (TP-IgM and TP-IgG) in human serum and investigated the expression of these antibodies during different stages of syphilis. METHODS: Serum samples were collected from 69 subjects (male 45, female 24) diagnosed with syphilis and analyzed by Western blotting for the expression of IgM and IgG against the four protein antigens. Expression levels of the target antibodies were compared during the same stage of syphilis as well as between different stages of this disease. RESULTS: In subjects with primary syphilis, the positive rate of Tp45 IgM was higher than that of other TP-IgM. Tp15 IgM was detected only in subjects with tertiary syphilis. Similarly, the seroprevalence of Tp45 IgG in primary syphilis was higher than for other TP-IgG. No target TP-IgM was detected in subjects with latent syphilis. In subjects with secondary syphilis, the expression level of Tp15 IgG (138.73 ± 20.16) was higher than for other target TP-IgG. In subjects with tertiary syphilis, all target TP-IgG were detected. In subjects with tertiary or latent syphilis, the expression levels of Tp45 IgG (121.33 ± 11.04 and 110.10 ± 40.19, respectively) were higher than those of other target TP-IgG. The expression levels of all Tp-IgM were similar before or after anti-syphilis treatment. In comparison, the expression levels of all TP-IgG decreased compared with the pre-treatment levels, and this decrease was statistically significant (both P < 0.05) for Tp17 IgG and Tp47 IgG. CONCLUSIONS: After Treponema pallidum infection, Tp45 IgM appeared first and Tp15 IgM occurred during later stages. The positive rates of all TP-IgG increased with the duration of this disease. Anti-syphilis treatment reduced the expression levels of Tp17 IgG and Tp47 IgG. Larger-scale studies are required to further validate the value of Tp15, Tp17, Tp45, and Tp47 as markers for the early detection of primary and latent syphilis.

Effects of Wei Chang An on expression of multiple genes in human gastric cancer grafted onto nude mice.

AIM: To investigate the expression of multiple genes in Chinese jianpi herbal recipe Wei Chang An (WCA) in human gastric cancer cell line SGC-7901. METHODS: A human gastric adenocarcinoma cell line SGC-7901 grafted onto nude mice was used as the animal model. The mice were randomly divided into 3 groups, one control and the two representing experimental conditions. Animals in the two experimental groups received either WCA over a 34-d period or 5-fluorouracil (5-FU) over 6-d period starting at 8th d after grafting. Control animals received saline on an identical schedule. Animals were killed 41 d after being grafted. The expression profiles in paired WCA treated gastric cancer samples and the N.S. control samples were studied by using a cDNA array representing 14181 cDNA clusters. The alterations in gene expression levels were confirmed by Real-time Quantitative polymerase chain reaction (qPCR). RESULTS: When compared with controls, the average tumor inhibitory rate in WCA group was 44.32% +/- 5.67% and 5-FU 47.04% +/- 11.33% (P < 0.01, respectively). The average labeling index (LI) for PCNA in WCA group and 5-FU group was significantly decreased compared with the control group. Apoptotic index (AI) was significantly increased to 9.72% +/- 4.51% using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method in WCA group compared with the controls 2.45% +/- 1.37%. 5-FU group was also found to have a significantly increased AI compared with the controls. The expression of cleaved Caspase-3 in WCA group and 5-FU group was significantly increased compared with the control group respectively. There were 45 different expressed sequence tags (ESTs) among the control sample pool and WCA sample pool. There were 24 ESTs up-regulated in WCA samples and 21 ESTs down-regulated. By using qPCR, the expression level of Stat3, rap2 interacting protein x (RIPX), regulator of differentiation 1 (ROD1) and Bcl-2 was lower in WCA group than that in control group respectively. By using SP immunohistochemical method the expression of Phospho-Stat3 (Tyr705) and Bcl-2 in WCA group and 5-FU group was significantly decreased compared with the control group respectively. CONCLUSION: WCA could inhibit gastric cancer cell SGC-7901 growth in vivo. WCA could induce gastric cancer cell apoptosis and suppress proliferation. Its mechanisms might be involved in the down-regulation of Stat3, RIPX, ROD1 and Bcl-2 gene.

Acoustic lens characterization for ultrasound and photoacoustic C-scan imaging modalities.

From a fundamental perspective, image reconstruction tasks in both ultrasound pulse echo and photoacoustic imaging are identical. We propose a C-scan imaging scheme that is applicable to both modalities where the image reconstruction is achieved through focusing action of an acoustic lens. The theory to characterize the imaging system is presented. Experimental methodology to determine the system point-spread-function is outlined and demonstrated with preliminary results.

[Difference of gene expression profile in human gastric cancer grafted onto nude mice treated with WCA].

OBJECTIVE: Chinese jianpi herbal recipe Weichangan (WCA) could increase the survival rate of advanced gastric cancer. This study was designed to investigate the molecular mechanism of WCA in treatment of gastric cancer by cDNA array, real-time quantitative PCR and immunohistochemical technique. METHOD: A human gastric adenocarcinoma cell line SGC-7901 grafted onto nude mouse was used as the animal model. The mice were divided into 3 groups, one control and the two representing experimental conditions. Animals in the two experimental groups received either WCA over a 34-day period or 5-fluorouracil (5-FU) over 6-day period starting at 8th day after grafting. Control animals received saline on an identical schedule. Animals were killed 41 days after being grafted. To assess the effect of therapy tumor weight was determined by a electron balance immediately after the animals killed. SP immunohistochemical method was used to detect the expression of proliferating cell nuclear antigen (PCNA) in xenografts. For detection of apoptotic cells, apoptotic indices (AI) were examined by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method. SP method was also used to detect the expression of cleaved Caspase-3. The expression profiles in paired WCA treated gastric cancer samples and the N. S. control samples were studied by using a cDNA array representing 14, 181 cDNA clusters. The alterations in gene expression levels were confirmed by real-time quantitative PCR. SP method was used to detect the expression of Phospho-Stat3 (Tyr705) and bcl-2. RESULT: When compared with controls, tumor growth was significantly inhibited by treatment with the WCA or 5-FU (P < 0.01, respectively). The average of tumor inhibitory rate in WCA group was (44.32 +/- 5.67)% and 5-FU (47.04 +/- 11.33)%. The average labeling index (LI) for PCNA in WCA group and 5-FU group was significantly decreased compared with the control group respectively. AI of human gastric cancer xenografts in nude mice was significantly increased to (9.72 +/- 4.51)% using TUNEL method in WCA group compared with the controls (2.45 +/- 1.37)%. 5-FU group was also found a significantly increased AI compared with the controls. The expression of cleaved Caspase-3 in WCA group and 5-FU group was significantly increased compared with the control group respectively. There were 45 different expression ESTs among the control sample pool and WCA sample pool. There were 24 ESTs up-regulated in WCA samples and 21 ESTs down-regulated. These 45 ESTs contains 35 cloned genes and 11 unknown ESTs. By using Real-time Quantitative PCR, the expression level of Stat3 (2(-deltadeltaCT) = 0.16) , RIPX (2(-deltadeltaCT) = 0.18), ROD1 (2(-deltadeltaCT) = 0.23) and bcl-2 (2 (-deltadeltaCT) = 0.10) was lower in WCA group than that in control group respectively. The expression of Phospho-Stat3 (Tyr705) and bcl-2 in WCA group and 5-FU group was significantly decreased compared with the control group respectively. CONCLUSION: Chinese jianpi herbal recipe WCA could inhibit gastric cancer cell SGC-7901 growth in vivo. WCA could induce gastric cancer cell apoptosis and suppress proliferation. Its mechanisms might be involved in the down-regulation of Stat3, RIPX, ROD1 and bcl-2 gene.

[Effects of Chinese herbal recipe Weichang'an in inducing apoptosis and related gene expression in human gastric cancer grafted onto nude mice].

OBJECTIVE: To investigate the mechanism of Chinese herbal recipe Weichang'an (WCA) in inducing cell apoptosis of human gastric cancer grafted onto nude mice. METHODS: The high performance liquid chromatography was used for monitoring the stability of WCA. A human gastric cancer cell line SGC-7901 grafted in nude mouse was used as the animal model. The mice were divided into untreated group and two experimental groups. Animals in the two experimental groups received either WCA over a 34-day period or 5-fluorouracil (5-FU) over a 6-day period starting at the 8th day after grafting. Animals in the untreated group received normal saline on an identical schedule. Animals were killed 41 days after being grafted. To assess the effect of the treatment on tumor, the tumor weight was determined by the electron balance immediately after the animals were killed. SP immunohistochemical method was used to detect the expression of proliferating cell nuclear antigen (PCNA) in grafts. Apoptotic indices (AI) of the tumor cells were examined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method. SP method was also used to detect the expressions of cleaved caspase-3, caspase-8 and caspase-9. SYBR green dye I real-time quantitative polymerase chain reaction (real-time quantitative [corrected] PCR) was used to assess the related gene alterations in mRNA level. The expressions of phospho-Stat3 (Tyr705) and bcl-2 proteins were detected by using SP method. RESULTS: Compared with the untreated group, tumor growth was significantly inhibited by treatment of WCA or 5-FU (P<0.01, respectively). The tumor inhibition rate in the WCA-treated group was 48.70% and that in the 5-FU-treated group was 60.10%. The average labeling index (LI) for PCNA in the WCA-treated group and 5-FU-treated group was significantly decreased as compared with that in the untreated group, respectively. The AI of human gastric cancer grafted in the nude mice detected by using TUNEL method was significantly increased to (9.72+/-4.51)% in the WCA-treated group, while it was (2.45+/-1.37)% in the untreated group. 5-FU-treated group was also found a significantly increased AI compared with the untreated group. The expressions of cleaved caspase-3 and caspase-9 in the WCA-treated group and 5-FU-treated group were significantly increased as compared with those in the untreated group. But caspase-8 showed no significant alteration either in the WCA-treated group or in the 5-FU-treated group. The expression levels of Stat3 (2(-)delta delta C(T))=0.16) and bcl-2 (2(-)delta delta C(T))=0.10) detected by using real-time quantitative [corrected] PCR were lower in the WCA-treated group than those in the untreated group. The expressions of phospho-Stat3 (Tyr705) and bcl-2 in the WCA-treated group were significantly decreased as compared with those in the untreated group. CONCLUSIONS: Chinese herbal recipe WCA can inhibit gastric cancer cell SGC-7901 growth in vivo, induce gastric cancer cell apoptosis and suppress the cell proliferation. WCA induces apoptosis through the caspase-9 and caspase-3 pathway in vivo. Its mechanism might be involved in the down-regulation of Stat3 and bcl-2 genes.

Difference of gene expression profile in human gastric cancer grafted onto nude mice treated with WCA / 中国中药杂志

<p><b>OBJECTIVE</b>Chinese jianpi herbal recipe Weichangan (WCA) could increase the survival rate of advanced gastric cancer. This study was designed to investigate the molecular mechanism of WCA in treatment of gastric cancer by cDNA array, real-time quantitative PCR and immunohistochemical technique.</p><p><b>METHOD</b>A human gastric adenocarcinoma cell line SGC-7901 grafted onto nude mouse was used as the animal model. The mice were divided into 3 groups, one control and the two representing experimental conditions. Animals in the two experimental groups received either WCA over a 34-day period or 5-fluorouracil (5-FU) over 6-day period starting at 8th day after grafting. Control animals received saline on an identical schedule. Animals were killed 41 days after being grafted. To assess the effect of therapy tumor weight was determined by a electron balance immediately after the animals killed. SP immunohistochemical method was used to detect the expression of proliferating cell nuclear antigen (PCNA) in xenografts. For detection of apoptotic cells, apoptotic indices (AI) were examined by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method. SP method was also used to detect the expression of cleaved Caspase-3. The expression profiles in paired WCA treated gastric cancer samples and the N. S. control samples were studied by using a cDNA array representing 14, 181 cDNA clusters. The alterations in gene expression levels were confirmed by real-time quantitative PCR. SP method was used to detect the expression of Phospho-Stat3 (Tyr705) and bcl-2.</p><p><b>RESULT</b>When compared with controls, tumor growth was significantly inhibited by treatment with the WCA or 5-FU (P < 0.01, respectively). The average of tumor inhibitory rate in WCA group was (44.32 +/- 5.67)% and 5-FU (47.04 +/- 11.33)%. The average labeling index (LI) for PCNA in WCA group and 5-FU group was significantly decreased compared with the control group respectively. AI of human gastric cancer xenografts in nude mice was significantly increased to (9.72 +/- 4.51)% using TUNEL method in WCA group compared with the controls (2.45 +/- 1.37)%. 5-FU group was also found a significantly increased AI compared with the controls. The expression of cleaved Caspase-3 in WCA group and 5-FU group was significantly increased compared with the control group respectively. There were 45 different expression ESTs among the control sample pool and WCA sample pool. There were 24 ESTs up-regulated in WCA samples and 21 ESTs down-regulated. These 45 ESTs contains 35 cloned genes and 11 unknown ESTs. By using Real-time Quantitative PCR, the expression level of Stat3 (2(-deltadeltaCT) = 0.16) , RIPX (2(-deltadeltaCT) = 0.18), ROD1 (2(-deltadeltaCT) = 0.23) and bcl-2 (2 (-deltadeltaCT) = 0.10) was lower in WCA group than that in control group respectively. The expression of Phospho-Stat3 (Tyr705) and bcl-2 in WCA group and 5-FU group was significantly decreased compared with the control group respectively.</p><p><b>CONCLUSION</b>Chinese jianpi herbal recipe WCA could inhibit gastric cancer cell SGC-7901 growth in vivo. WCA could induce gastric cancer cell apoptosis and suppress proliferation. Its mechanisms might be involved in the down-regulation of Stat3, RIPX, ROD1 and bcl-2 gene.</p>

[Growth-inhibiting and anti-metastasis effects of Weichang'an Decoction on orthotopic transplant nude mouse model of human gastric cancer].

OBJECTIVE: To evaluate the growth-inhibiting and anti-metastasis effects of Weichang'an Decoction (WCAD) on orthotopic transplant nude mouse model of human gastric cancer. METHODS: Forty-one nude mice were implanted with SGC-7901 cells at orthotopic site, whereas 25 were implanted with SGC-7901 cells subcutaneously. Then the nude mice in each transplantation model were divided into the same three groups which were WCAD-treated group with WCAD 0.5 ml/d taken orally, 5-FU-treated group with 5-FU 50 mg/kg intraperitoneally injected weekly and untreated control group with physiological saline 0.5 ml/d taken orally. The growth-inhibiting rates of transplant tumors were detected and the metastatic lesions were examined in the orthotopic transplant mouse model while PCNA-positive rate and apoptotic index (AI) were observed in the subcutaneous transplant mouse model. RESULTS: The growth-inhibiting rates in the WCAD-treated and 5-FU-treated groups of orthotopic transplant mouse model were 40.82% and 37.92% respectively whereas those of subcutaneous transplant mouse model were 48.70% and 60.10%. The incidence rates of metastasis in perigastric lymph notes, lymph nodes in the porta hepatis, liver, diaphragm and peritoneum in the WCAD-treated and 5-FU-treated groups were lower than those in the untreated control group, and the total metastasis rates in the WCAD-treated, 5-FU-treated and the untreated control groups were 30.77%, 28.57% and 71.43% respectively with significant differences (P<0.05). The total PCNA-positive rates in the WCAD-treated and 5-FU-treated groups were obviously lower than that in the untreated control group (P<0.05 or P<0.01) while the AI was higher than that in the untreated control group (P<0.01). The growth-inhibiting rate, total PCNA-positive rate and total metastasis rate in the WCAD-treated group had no significant differences as compared with those in the 5-FU-treated group, but the AI in the WCAD-treated group was significantly higher than that in the 5-FU-treated group (P<0.05). CONCLUSION: WCAD has the inhibiting effects on tumor growth and metastasis of gastric cancer which is orthotopic implanted onto nude mice. This effect may be obtained by proliferation suppression and apoptosis induction in cancer.

Effects of Chinese Jianpi herbs on cell apoptosis and related gene expression in human gastric cancer grafted onto nude mice.

AIM: To explore the mechanism of the Sijunzi decoction and another Chinese herbal recipe (SRRS) based mainly on the Sijunzi decoction in treatment of gastric cancer. METHODS: A human gastric adenocarcinoma cell line SGC-7901 grafted onto nude mouse was used as the animal model. The mice were divided into 3 groups, one control and the two representative experimental conditions. Animals in the two experimental groups received either Sijunzi decoction or SRRS over a 40-day period starting at 1st day after grafting. Control animals received saline on an identical schedule. Animals were killed 41 days after being grafted. The effect of therapy was assessed by two ways: (1) tumor size was periodically measured during the life of the animals; (2) tumor weight was determined by a electron balance immediately after the animals killed. For detection of apoptotic cells, apoptotic indices(AI) were examined by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method. Morphological alterations were observed with electron microscopy. S-P immunohistochemical method was used to detect the expression of Ki-67 in xenografts. Expression of bcl-2 and p53 was semiquantitatively detected using a reverse transcriptase-polymerase chain reaction (RT-PCR) technique. RESULTS: When compared with controls, tumor growth (size and weight) was significantly inhibited by treatment with the Sijunzi decoction (P<0.05) or SRRS (P<0.01). The tumor inhibitory rate in the Sijunzi decoction group was 34.33 % and SRRS group 46.53 %. AI of human gastric cancer xenografts in nude mice was significantly increased to 16.24+/-3.21 % using TUNEL method and 11.38+/-6.46 % by FACScan in the Sijunzi decoction group compared with the controls (TUNEL: 2.63+/-1.03 %, P<0.01; FACScan: 7.15+/- 1.32 %, P<0.05). SRRS group was also found a significantly increased AI by using TUNEL method and flow cytometry analysis compared with the controls (TUNEL: 13.18+/-3.05 %, P<0.05; FACScan: 11.58+/-5.71 % (P<0.05). Under electron microscope, cell shrinkage, nuclear chromatin condensation, formation of membrane blebs and apoptotic bodies were frequently observed in Sijunzi decoction group and SRRS group. The average labeling index (LI) for Ki-67 in SRRS group was significantly decreased to 8.43+/-2.22 % compared with the control group (10.37+/-4.91 %) (P<0.05). The average labeling index for Ki-67 in sijunzi decoction group was 7.95+/- 2.54 % which was lower than that of the control group, but showed no significance (P=0.07). The expression level of p53 mRNA was lower in both Sijunzi decoction group and SRRS group than that in control group (P<0.05; P<0.01). The expression of bcl-2 mRNA was also decreased in SRRS group compared with the control (P<0.01). CONCLUSION: The inhibition of gastric cancer cell growth in vivo by Chinese Jianpi herbs and SRRS is related to induction of the cell apoptosis which may be involved in aberrant expression of p53 and bcl-2 genes.